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    18 December 2022, Volume 39 Issue 6 Previous Issue    Next Issue
    Inhibition and mechanism of AEAMP9, a novel compound formula of alcoholic extract of anticancer medicinal plants,on lung cancer A549 cells
    AO Zhiguang, JIANG Yuhong, YANG Lei, LI Dong, WANG Jun, WANG Siyi, MAO Canquan
    2022, 39 (6):  1.  doi: 10.3969/j.issn.2095-1736.2022.06.001
    Abstract ( 266 )   PDF   Save
    In view of the high morbidity and mortality of lung cancer, lung cancer was concerned and human non-small cell lung cancer A549 cells were used as the target in this study. Here, after screening 124 kinds of medicinal plants in A549 lung cancer cells and the determination of their anticancer orders, an efficient and novel compound formula of AEAMP9 was developed and its underlying molecular mechanism was investigated. AEAMP9 was prepared from the alcoholic extracts of six anticancer medicinal plants including Hedyotis diffusa Willd. (Bai Hua She She Cao), Phellodendron amurense Rupr. (Huang Bai), Glycyrrhiza uralensis Fisch. (Gan Cao), Platycladus orientalis (L.) Franco (Ce Bai Ye), Chrysanthemum indicum L. (Ye Ju Hua) and Notopterygium incisum Ting ex H. T. Chang (Qiang Huo). It can effectively inhibit the viability and clone formation of cancer cells, provoke apoptosis and block migration in A549 cells. Mechanistically, AEAMP9 significantly decreased the expressions of ABCG2, FoxM1, Vimentin, MMP14 and the ratio of Bcl-2/Bax, consequently promoted the activation of apoptosis associated genes including p53, caspase 3, caspase 8 and caspase 9. Network pharmacology and transcriptome RNA-seq revealed that many genes were significantly and differentially expressed in A549 cells after treatment with AEAMP9. TNF, IL-17 and other important signal pathways related to tumor, inflammation and apoptosis were closely regulated. EGR1, IL-6, ALK and VEGF may be the key regulatory targets for the interaction of AEAMP9 and lung adenocarcinoma. The components of quercetin, kaempferol and luteolin in AEAMP9 may play an important role in antitumor activity. Safely, AEAMP9 had no obvious toxic effects on SPF Kunming mice. Furthermore, AEAMP9 can significantly inhibit the tumor growth in mouse A549 xenograft models
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    Mechanism of curculigoside against PTSD-induced inflammatory response in mouse hippocampus
    JI Manman, ZHANG Zhengrong, ZHU Guoqi
    2022, 39 (6):  14.  doi: 10.3969/j.issn.2095-1736.2022.06.014
    Abstract ( 220 )   PDF   Save
    Curculigoside (CUR) is the main component of Curculigo and has various biological activities. However, it is not clear whether CUR inhibits post-traumatic stress disorder (PTSD)-induced inflammation in the hippocampus of mice. In this study, a modified mouse model of single prolonged stress and electrical stimulation (SPS&S) was established to investigate the effect of curculigoside on neuroinflammation in the hippocampus of SPS&S-treated mice from the prospective of nuclear factor kappa-B (NF-κB) pathway. The results showed that, compared with that of SPS&S group, CUR significantly reduced the freezing time of PTSD mice during fear extinction (P<0.05). It increased the movement time and distance in the central area in the open field test (P<0.05), and increased the number of open arms in the Elevated Plus Maze Test (P<0.05). In addition, compared with that of SPS&S group, CUR decreased the levels of TNF-α and IL-1β in serum of PTSD mice (P<0.05). It improved the pathological changes in CA1 regions of the hippocampus, and decreased hippocampal expressions of NF-κB, NLRP3, and Caspase-1(P<0.05). These data implicated that CUR improved PTSD-like behaviors in mice, which might be related to the regulation of NF-κB-dependent neuroinflammation.
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    Recombinant expression and purification of the extracellular domain of human B lymphocyte surface antigen
    CHANG Qing, YU Minda, LI Wenqi
    2022, 39 (6):  20.  doi: 10.3969/j.issn.2095-1736.2022.06.020
    Abstract ( 175 )   PDF   Save
    Human CD20 is a B lymphocyte surface antigen abnormally high-expressed in B-cell lymphoma, chosen as a cell-therapy target of B-cell malignancies. By molecular cloning, the extracellular domain of human CD20 was successfully constructed into pET28a vector as twins connected with a linker (-GSSGGSSG-), and with a C-terminal 6His-tag. After confirming by DNA sequencing, the expression vector named pET28a-Bi20 was transformed into E. coli Transetta(DE3). The expression of the recombinant product Bi20 was induced by adding 1 mmol/L IPTG. When the optical density at 600 nm (OD600) of the bacterial suspension reached 0.6-0.8, at 37 ℃ for 5 h. The recombinant Bi20 was expressed as inclusion body. Through washing of inclusion body, purifying by Ni-affinity column under denatured condition, and screening and optimizing the refolding experimental conditions of inclusion body, the target peptide was finally purified as a soluble form of high purity. Western blotting and ELISA demonstrated that the purified Bi20 protein had a desirable immunogenicity.
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    Purification and characterization of F-actin depolymerization properties of cofilin from Neurospora crassa
    HUANG Lu, CHEN Jingqiu, SUN Haitao, LI Yanhong, CHEN Zhiling
    2022, 39 (6):  25.  doi: 10.3969/j.issn.2095-1736.2022.06.025
    Abstract ( 142 )   PDF   Save
    In order to clarify biochemical properties of cofilin from Neurospora crassa (NcCof), purification and characterization of F-actin depolymerization properties of NcCof were performed in this study. Sequences alignment indicated that Ser4 in the N-terminus of NcCof was highly conserved and might be important for its F-actin severing activity. Three-dimensional structure analysis showed that NcCof was composed of five α-helices and six β-strands. Of them, four internal β-strands were antiparallel, and all the β-strands were surrounded by the five α-helices,which were consistent with the typical structure of cofilin/ADF family. Subsequently, the S4 of NcCof was performed site-directed mutagenesis and two proteins NcCof(S4A)and NcCof(S4D)were obtained after purification. Furthermore, high-speed co-sedimentation assay was used to determine and compare the F-actin depolymerizing activity. The result showed that the control (NcCof) demonstrated the intrinsic and concentration dependent F-actiin depolymerizing activity, while this activity of both NcCof(S4A) and NcCof(S4D) displayed a weakening trend, especially the F-actin depolymerizing activity of NcCof (S4D) decreased significantly. This research conformed that S4 of NcCof was the key site for the F-actin depolymerization in Neurospora crassa, which would be helpful in further exploring the mechanisms of regulating actin dynamics by NcCof.
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    The effect of interfering VDR on HepG2 cell proliferation
    CHENG Jia, JI Zhengwei, WU Xifeng, LU Hongzhao, ZHANG Tao
    2022, 39 (6):  30.  doi: 10.3969/j.issn.2095-1736.2022.06.030
    Abstract ( 134 )   PDF   Save
    To investigate the effect of vitamin D receptor (VDR) in HepG2 cell proliferation and the significance of VDR in liver cancer, HepG2 cells were transfected with VDR gene-specific siRNA (small interfering RNA). The qPCR and WB results showed that compared with the control NC siRNA group, VDR siRNA-507 could significantly inhibit the mRNA and protein levels of VDR (P<0.05). The celltiter-Lumi luminescence, EDU staining and cell colony formation assay were employed to detect the effect of interference VDR on the proliferation of HepG2 cells. The results confirmed that interference with VDR could significantly promote the proliferation of HepG2 cells (P<0.01). The immunofluorescence and double luciferase activity assay were used to monitor the RBP4 expression during HepG2 cells growth process. However, interfering VDR had no effect on RBP4 expression level in HepG2. It was concluded that interference with VDR could promote HepG2 cells proliferation which was no relation with RBP4 level.
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    Predictive study on the potential molecular mechanism of SARS-CoV-2 spike protein RBD mutation to enhance its infectivity
    LIU Juzhao, YANG Yuping, XU Jianbo, LYU Hongchang, CHEN Junyu, ZHANG Chunchun, CUI Qi
    2022, 39 (6):  35.  doi: 10.3969/j.issn.2095-1736.2022.06.035
    Abstract ( 319 )   PDF   Save
    After the outbreak of SARS-CoV-2, more and more mutants with stronger infectivity appeared. The most representative strains were B.1.351 and B.1.617, which had a common feature of amino acid mutation at spike protein 484 site. Site 484 is a site where interacts with residue K31 on hACE2. From the perspective of molecular dynamics and protein docking, E484K and E484Q mutations of SARS-CoV-2 were investigated, and it was found that the hydrogen bonds between amino acid residues (A475 and N489) that directly interact with hACE2 were shortened due to 484 mutations. Kinetic results also showed that amino acid sequence 475-489 was highly active and should not be neglected as drug design targets. It can provide a new target for the development of SARS-CoV-2 inhibitors, predict the future evolutionary direction of SARS-CoV-2 to a certain extent, and also provide a new idea for the mutation research of similar coronavirus.
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    Functional analysis of a C6-type transcription factor Cgctf1 in Colletotrichum gloeosporioides
    CAO Jian, LIU Shayu, WANG Diguang, LIU Zhiqiang
    2022, 39 (6):  41.  doi: 10.3969/j.issn.2095-1736.2022.06.041
    Abstract ( 241 )   PDF   Save
    C6-type transcription factor plays an important role in regulating the growth, development and pathogenicity of plant pathogenic fungi. A C6-type transcription factor gene Cgctf1 was identified from C. gloeosporioides, which encodes 405 amino acids and contains a GAL4 domain. Gene-knockout mutants of Cgctf1 were obtained by homologous recombination, and the complementary strain was obtained on the basis of the mutants. Phenotypic analysis showed that there was no significant difference in vegetative growth between the knockout mutant and the wild type, but the mutant was more sensitive to salt stress and more resistant to H2O2 and SDS. The conidia yield, spore germination rate and appressorium formation rate of the knockout mutant were significantly lower than those of the wild type. Onion epidermis infection and cellophane invasion tests showed that the penetration ability of conidia of the knockout mutant was weakened. On the rubber leaves, the disease spot formed by knockout mutants was smaller than that of wild type, and the mutants had reduced virulence. These results suggest that Cgctf1 is involved in the regulation of conidial development, appressorium formation and invasion, stress response and pathogenicity of C. gloeosporioides.
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    Homologous overexpression of laccase Lcc9 in Coprinopsis cinerea
    ZHANG Jingna, ZHOU Gang, LIU Juanjuan, FANG Zemin, XIAO Yazhong
    2022, 39 (6):  47.  doi: 10.3969/j.issn.2095-1736.2022.06.047
    Abstract ( 184 )   PDF   Save
    In order to prepare a large amount of Lcc9, the gene lcc9 was cloned and homologously overexpressed in Corprinopsis cinerea Okayama 7#130 (OK7) and FA2222. A total of 46 OK7-lcc9 and 29 FA2222-lcc9 positive transformants were obtained, respectively, based on the primary screening in regeneration medium and re-screening in minimal medium. After inoculation of 5% (V/V) seed cultures in Kjalke medium and cultured at 37 ℃ for 6-7 d, laccase activity of OK7-lcc9 and FA2222-lcc9 in shaking flasks reached 5.1-21.1 U/mL and 3.7-12.3U/mL, respectively, 19.2 and 241.2 times higher than that of the wild type strains. One of the FA2222 positive strains was cultured in a 3 L fermenter for laccase preparation. Results showed that the laccase activity of the strain reached 59 U/mL after incubation at 37 ℃ and 50 r/min for 108 h, which was 4.8 times higher than that in the liquid shake flask cultures.
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    Optimization of cultivation conditions of a non-specific nuclease from Yersinia enterocolitica subsp. Palearctica
    ZHU Yu, GUO Senlin, GAO Ting, WANG Lu, LI Duanhua, LI Jinjun, GE Yan, ZHAO Chen
    2022, 39 (6):  52.  doi: 10.3969/j.issn.2095-1736.2022.06.052
    Abstract ( 144 )   PDF   Save
    To obtain the high-level extracellular expression of a non-specific nuclease from Yersinia enterocolitica subsp. palearctica (Nucyep), the gene fragments of nucyep from pET-24a-nucyep were cloned into the shuttle vector pBE-S to yield pBE-S-nucyep. The linearized pBE-S-nucyep was ligated to the DNA mixtures of different signal peptides through In-Fusion cloning enzyme, then transformed into competent E. coli Stellar cells. 43 positive clones were obtained and the corresponding recombinant plasmids pBE-S-nucyep-SP were separately transformed into Bacillus subtilis (B. subtilis) RIK1285 to yield 43 recombinant strains. The Nucyep activities of 43 recombinant strains were determined using the specific optical density method which has been developed in our lab previously. B. subtilis RIK1285/pBE-S-nucyep-YoaW carrying the signal peptide YoaW showed the highest enzymatic activity level among the engineered strains and was employed in the next study. Through optimization of cultivation conditions, the highest Nucyep activity of (8.27±0.41) U/μL was obtained and it exhibited 20-fold higher than that of the cultivation conditions before optimization. The optimized cultivation conditions were as follows: volume fraction 5% of seed culture inoculum, culture medium 3×LB, incubated at 30 ℃, 48 h and 220 r/min. In conclusion, the results indicated that the high activity of Nucyep could be achieved through the employment of suitable signal peptide in combination with applying the optimized cultivation conditions. It will provide some support for further study.
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    Effects of yeast fermentation on bioactivity and active components of Paeonia lactiflora Pall. pods #br#
    CAO Qingchao, LU Yuting, JIN Yinzhe, LIN Jianxing, TAO Jun , JIN Yingshan
    2022, 39 (6):  59.  doi: 10.3969/j.issn.2095-1736.2022.06.059
    Abstract ( 130 )   PDF   Save
    In order to fully exploit and utilize of by-products of P. lactiflora Pall. for oil, the yeast fermentation of P. lactiflora Pall. pods (PLPPs) was carried out by microbial fermentation technology to evaluate the changes of fermentation on bioactivity and active components of PLPPs, and the molecular docking simulation was used to analyze the affinity constants between the active components of PLPPs and the main anti-aging enzymes in vivo. The results showed that the total polyphenols content of the Saccharomycopsis fibuligera (SF) fermentation product (SF-F) of PLPPs was the highest (338.29±0.99) mg GA/g DW, which was increased by 23.49% compared with that of non-fermentation (N-F). The total antioxidant activity, total reducing ability and acetylcholinesterase inhibition on SF-F were the highest, which were 38.11%, 31.89% and 107.24% higher than those of N-F, respectively. LC-MS analysis showed that the content of gallic acid in PLPPs was significantly increased after being fermented by SF, while 1,2,3,4,6-O-pentagalanyl-β-D-glucose, quercetin-3-O-β-D-glucose-7-O-β-D-gentiandiglycoside, quercetin-3-O-glucoside, and 2″-O-galacylhyperoside disappeared. Molecular docking simulation results showed that the main active components of PLPPs had a certain docking affinity with anti-aging enzymes. The results indicated that some macromolecular compounds in PLPPs were transformed into small molecular compounds by SF fermentation, which increased the total polyphenols content and bioactivity after fermentation.
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    Identification of chromosome ploidy in kiwifruit germplasm by flow cytometer
    LUO Qing, GAO Jianyou, LI Jiewei, YE Kaiyu, MO Quanhui, JIANG Qiaosheng, ZHA Manrong, WANG Faming, LIU Shibiao
    2022, 39 (6):  66.  doi: 10.3969/j.issn.2095-1736.2022.06.066
    Abstract ( 228 )   PDF   Save
    Ploidy identification is one of the preconditions for kiwifruit breeding. In this study, 50 kiwifruit germplasms were collected from Guilin, Guangxi Province, and their ploidy was identified using flow cytometry. Among them, the ploidy of 16 germplasms was identified for the first time, and the relationship between ploidy and fruit main features was analyzed. The results showed that 19 diploid, 13 tetraploid and 18 hexaploid were identified from the 50 species and varieties, the majority of Actinidia Chinensis were diploids and tetraploids, with red or yellow-fleshed and smooth or short trichomes on the fruit surface, while that of A.deliciosa was hexaploidy, with generally green-fleshed and long velutinous. The fruit size of A.deliciosa was larger than that of A. chinensis totally. Wild species were mostly diploids, accompanied with green-fleshed and smooth or less trichomes. This study enriched the ploidy database, and provided references for breeding and production of kiwifruit.
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    Chromosome atlas of three species from the Qinghai-Tibet Plateau
    MA Xiaolei, LIU Ruijuan, CAO Qian, XING Rui, CHEN Shilong, GAO Qingbo
    2022, 39 (6):  73.  doi: 10.3969/j.issn.2095-1736.2022.06.073
    Abstract ( 141 )   PDF   Save
    The seeds of three endemic plants collected from the Qinghai-Tibet Plateau and germinated on wet filter papers and root tip-squash method were used for cytological observation.The results showed that three species were diploid. Chromosome numbers of all 5 populations of Micranthes melanocentra were 2n=22; chromosome numbers of 2 populations of Micranthes atrata were 2n=22. All 5 populations of Swertia przewalskii had same chromosome numbers, the karyotype formula was 2n=24=20m+4sm, 1B,As. K%=58.8%. The third and ninth pairs were submetacentric, and the rest were metacentric. The chromosome cardinality of Micranthes melanocentra was consistent with previous studies, but no polyploid was found, indicating that the polyploid frequency was low in the sampling area. The karyotypes of Micranthes atrata and Swertia przewalskii were reported for the first time. The reported karyotypes of the three plants complemented the cytological data of the species endemic to the Qinghai-Tibet Plateau, and provided cytological data for the development, utilization and conservation of these species.
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    Transcriptomic analysis of Dunaliella polymorpha under different light intensity stresses #br# #br#
    GAO Fan, YANG Peng, WANG Fei, XU Lu, TANG Liqun
    2022, 39 (6):  77.  doi: 10.3969/j.issn.2095-1736.2022.06.077
    Abstract ( 146 )   PDF   Save
    In order to research the response mechanism of Dunaliella polymorpha under different light stresses, screen differentially expressed genes, and analyze their functions and metabolic pathways, the shading control group, medium light intensity stress group and high light intensity stress group were set up respectively based on the mature algae strains. The analyses were performed by utilizing morphological, fluorescence parameter determination, high-throughput sequencing and bioinformatics methods at the cellular, biochemical and transcriptomic levels, respectively.A total of 52853 unigenes with 29025 coding domain sequences (CDSs) from the nine RNA-seq libraries were obtained in this alga. 41.59% of unigenes were annotated in the 7 bio-database. Moreover, a total of 282 contigs were de novo assembled with a 1072 bp N50 and a GC percentage of 52.25%.Furthermore, based on the differentially expressed genes (DEGs), the medium light intensity stress group and high light intensity stress group could be clusted together and the number of down-regulated DEGs (9385) was more than that of the up-regulated ones (8046). The GO and KEGG pathway analyses of the intergroup common DEGs showed that the functions of key genes involved in the light stress response were mainly related to the intracellular photosynthetic catalytic activity and antenna protein synthesis. The main metabolic pathways were related to the metabolic process stimulated by the light intensity. The results laid a foundation for further researching on this algal photosynthetic mechanism, cloning and expression of key photosynthetic regulation genes, molecular breeding and their industrial application in future.
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    Effects of different ways of adding 4th urease inhibitors into urea on the growth and some quality indexes of Chinese cabbage
    WANG Chenyi, DONG Lei, ZHANG Xueyi, ZHAO Yuhan, HUANG Zhaowei
    2022, 39 (6):  84.  doi: 10.3969/j.issn.2095-1736.2022.06.084
    Abstract ( 148 )   PDF   Save
    In order to study the effects of the different ways of adding 4th urease inhibitor (4th UI) into urea on the growth and some quality indexes of Chinese cabbage, parameters such as the fresh weight, plant height, leaf length, leaf width, chlorophyll content, soluble sugar and root activity of Chinese cabbage were studied by outdoor potting method in this paper. The results showed that the yield, quality index and nutrient index of Chinese cabbage could be improved under three different ways and dosage of adding inhibitors into urea. Among them, 1% of the inhibitor concentration in group B combined with urea had the best yield increasing effect, reaching 32.82%. 0.2% of the inhibitor concentration in group C had the best effect, with yield increasing by 80.49%, plant height increasing by 17.34%, total nitrogen content of plants increasing by 61.36%, and total phosphorus content of plants increasing by 78.85%, the root activity of cabbage increasing by 26.61%. In group D, the best effect was 50.01% when 0.2% urease inhibitor was added into urea. In the same method of administration, the urease inhibitors with low and medium concentration had the best effect on increasing yield of crops. Comprehensive consideration, the group C with adding 0.2% inhibitors into urea was the best way. The results can provide a basis for the application of this new type of slow-release urea in agriculture.
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    Research progress on anti-tumor effect of natural product active molecule erianin
    HONG Jing, YANG Fangyao, JIANG Lixiang, JIAN Tiantian, WANG Siyu, HUANG Xinhe
    2022, 39 (6):  89.  doi: 10.3969/j.issn.2095-1736.2022.06.089
    Abstract ( 211 )   PDF   Save
    Natural products are an important source of anti-tumor drugs. Based on the sorting and analysis of current studies at home and abroad, this paper summarized the anti-tumor effect and anti-tumor molecular mechanism of bibenzyl small molecule erianin in medicinal plants Dendrobium candidum and Dendrobium chrysotoxum. Firstly, the medicinal history and pharmacological activities of Dendrobium natural products, as well as the various pharmacological effects of erianin, especially its antitumor effects were introduced and explored. Combined with a number of in vitro and in vivo studies, the inhibitory effects of erianin on tumor proliferation of many cancer cell lines were introduced. By combing and analyzing the available research, this paper analyzed the role of erianin in tumor regulation by inducing cancer cell apoptosis, autophagy, blocking cancer cell cycle, inhibiting cancer cell migration and invasion, inhibiting tumor angiogenesis and other molecular mechanisms, and summarized several erianin related signal pathways and factors such as JNK, PI3K / Akt, ERK. Finally, in view of research status of erianin. The prospect of erianin in patent medicine research and clinical application was prospected.
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    Research progress on glycoside hydrolases gene family of plant pathogen
    PAN Fengying, LIU Lulu, SUN Dayun, GUO Zexi, QU Junjie, YIN Ling
    2022, 39 (6):  94.  doi: 10.3969/j.issn.2095-1736.2022.06.094
    Abstract ( 380 )   PDF   Save
    This paper reviews the research progress of plant pathogen glycoside hydrolases (GHs) gene family in several aspects, including classification, enzyme activity, distribution characteristics and roles in pathogen infection. GHs’ roles in pathogen infection were focused. GHs play the role of virulence factors/pathogenic factors promoting pathogen infection and the role of elicitors or pathogen-associated molecular patterns (PAMPs) inducing plant immunity. GHs decompose plant cell walls into damage-associated molecular patterns (DAMPs) to induce plant immunity and the specific binding or breakdown pathogen/microbe-associated molecular patterns (P/MAMPs) to suppress plant immunity. On these bases, the molecular mechanism of plant pathogen GHs regulating plant immunity and existing problems were summarized.It provided reference for in-depth studies of more functions of GHs, as well as for better utilization of these GHs family members in crop improvement and disease control strategies.
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    New application of overlapping genes in the expression of heterologous proteins
    PENG Kai, LU Xiaoyun, LU Lina, ZHANG Juankun, JIANG Huifeng
    2022, 39 (6):  101.  doi: 10.3969/j.issn.2095-1736.2022.06.101
    Abstract ( 183 )   PDF   Save
    There are still difficulties in regulating the expression and solubility of heterologous proteins in E. coli. In this study, a protein expression optimization method that combines overlapping gene structure and 5′-end sequence optimization of mRNA was developed. Utilizing the 5′-end tag of the pET-28a plasmid vector expression box, and the target gene introduced into TG before the start codon formed an overlapping gene type expression structure of TGATG. Then, by designing degenerate primers, synonymous codon mutations of 14 consecutive codons at the 5′-end of a heterologous gene mRNA were performed to change the secondary structure of mRNA and realize the screening of overlapping gene structure strength. This method can affect the expression of heterologous protein without changing the plasmid composition and external conditions, and regulate the solubility of the protein by affecting the rate of the translation process. Several examples of heterologous gene expression optimization in this study have proved the role of overlapping gene structures in regulating the soluble expression of proteins. It provided a simple optimization strategy for the regulation of the expression and solubility of heterologous proteins in E. coli.
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    Application of βGlcY dyeing technology in histochemistry localization of arabinogalactan proteins in Ziziphus jujuba Mill cv. Lingwuchangzao
    ZHANG Yingcai, WANG Jing, TAO Shanshan
    2022, 39 (6):  108.  doi: 10.3969/j.issn.2095-1736.2022.06.108
    Abstract ( 138 )   PDF   Save
    The different developmental stages fruits of Ziziphus jujuba Mill cv. Lingwuchangzao were used as experimental material to study the distribution of arabinogalactan proteins(AGPs) by βGlcY dyeing technology and confirmed by immunofluorescence localization. The main research results were followed: the brown-red precipitate formed by βGlcY-AGPs distributed in the cell walls and cell interiors of the epicarp and the alongside internal layers of closely arranged mesocarp small cells in different periods fruit. The brown-red precipitate formed by βGlcY-AGPs distributed in the cell walls and the inner part of the large ovoid parenchyma cells in the inner mesocarp during the early bulking period, while they were mainly distributed in the parenchyma cell walls during the other periods, with no distribution in the inner part of most cells. The brown-red precipitate formed by βGlcY-AGPs distributed in the cell walls and cell interiors of all cells in the vascular bundles in different periods fruit, the distribution of brown-red precipitate formed by βGlcY-AGPs gradually decreased in the mesocarp parenchyma cells with fruit development, this confirmed that AGPs gradually decreased. The intensity of brown-red precipitate formed by different concentration βGlcY was different at the same periods, the higher the concentration, the better the dyeing effect. There were some differences in brown-red precipitate intensity formed by the same concentration βGlcY in different tissues of fruit at different periods. βGlcY dyeing technology was effective method for AGPs histochemical localization of Ziziphus jujuba Mill cv. Lingwuchangzao fruit.
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    Reform and innovation of genetics basic experimental teaching in universities
    GUO Yan, WANG Naining, ZHU Dongli, YANG Tielin
    2022, 39 (6):  114.  doi: 10.3969/j.issn.2095-1736.2022.06.114
    Abstract ( 189 )   PDF   Save
    Under the current background of cultivating innovative, comprehensive and compound talents, the proportion of basic verification experiments in molecular genetics experiments in the curriculum of biology majors is becoming less and less. However, the lower foundation determines the superstructure. Our university has carried out corresponding teaching reforms and innovations on the molecular genetics genotype verification experiment in the genetic experiment course of biotechnology, and added curriculum ideology and politics elements of the course,aiming to integrate the elements of cultivating students’ scientific thinking and innovation ability into the basic verification course, so as to cultivate students’ interest in learning, mobilize the enthusiasm for class, improve students’ innovative thinking ability and practical ability, and lay a foundation for the selection and training of top-notch innovative talents in basic disciplines of the “Plan for strengthening basic disciplines”.
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    Diagram reading-based evaluation of biology professional abilities
    MA Xiaoyan, CHEN Zhenya, HUO Yixin
    2022, 39 (6):  117.  doi: 10.3969/j.issn.2095-1736.2022.06.117
    Abstract ( 174 )   PDF   Save
    This paper proposes a type of test question based on diagram interpretation. By interpreting relevant diagrams of theoretical concepts, experimental data, engineering strategies, etc., the biological process could be explained, theoretical questions could be answered, and engineering solutions could be designed, thus realizing a comprehensive examination of the three key abilities of biology majors such as information extraction, logical thinking and theoretical application. This paper analyzed the characteristics and advantages of the diagram-reading questions with representative cases, and the design principles of such question were proposed. Implementation of the diagram-reading questions showed that such question was significantly different from the traditional memory-based questions, which could effectively eliminate the "memory type" candidates, exhibit excellent score distinction and selection accuracy.Such diagram-reading question provided a novel method for the evaluation of actual professional ability. It promoted the upgrading and optimization of the selection method of biological professionals, and guarantees the quality of higher biology education.
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    Exploration on the reform of experimental examination of parasitology based on the national parasitic resource center of China
    ZOU Jiexin, ZHANG Liyanyang, SHEN Haimo, ZHU Chunchao, SHI Linbo, ZHANG Meng, GAO Dian, ZHANG Xiaoyan
    2022, 39 (6):  122.  doi: 10.3969/j.issn.2095-1736.2022.06.122
    Abstract ( 184 )   PDF   Save
    In the study of parasitology, it is very important to master the morphological characteristics of various parasites for the diagnosis of diseases. The way of observing physical specimens is usually adopted in the traditional parasitology experimental examination. However, this examination method has many limitations. After the emergence of COVID-19, it is urgent to establish an online examination system for parasitology. In this study, we have obtained the pictures of various parasite specimens collected in the national parasitic resource center (the primary sub platform of the national science and technology resource sharing service platform) in real time. An online examination system based on B/S architecture was built on the internet and applied to experimental teaching of parasitology. The system has the functions of random generation of test paper, online examination, intelligent marking and so on. The examination form is novel and easy to operate, which improves the feasibility, impartiality and credibility of the examination.
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    Comprehensive experimental development and evaluation of physiological mechanism of non-gravity bending of primary roots of legumes
    MA Shaoying, MA Lei, TIAN Peng, YANG Jie, LI Sheng, WANG Tingting
    2022, 39 (6):  127.  doi: 10.3969/j.issn.2095-1736.2022.06.127
    Abstract ( 149 )   PDF   Save
    Some contents of the plant physiology experiment large course for postgraduates were reformed in order to cultivate students’ interest in scientific research, establish scientific research thinking, and improve teaching quality. A comprehensive experiment of physiological mechanism of non-gravity bending of legume seedling roots was designed. The findings revealed that by consulting the literature, engaging in a discussion with teachers and students, reforming the experimental scheme and incorporating the experimental content, and other similar activities. The students’ interest in learning was stimulated, their awareness of innovation and exploration was established, and their ability to conduct scientific research was improved. This suggested that the development of comprehensive experiments was more beneficial to students in understanding scientific problems by integrating all knowledge points than a traditional experiment and in carrying out the experiment relative to obtaining dissertations. It could achieve good teaching effect and provide a consult for carrying out more comprehensive experiments in the future.
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