生物学杂志
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摘要： 通过紫外诱变筛选出的高产洛伐他汀红曲霉菌株M120-1为出发菌株，提取红曲霉菌株总RNA ，通过RT-PCR技术克隆LovD基因，构建表达载体，对红曲霉进行遗传转化；通过高效液相色谱法筛选高产洛伐他汀转化菌株。结果表明：提取红曲霉总RNA并建立表达载体，随机挑选出12个转化子，经PCR鉴定，有10株成功得到转化。对其进行培养并通过HPLC法测定洛伐他汀产量，其中一株转化菌株的洛伐他汀产量为0.55 mg/mL，比出发菌株产量高63%。对该菌株进行传代培养，具有良好的遗传稳定性。
关键词: 红曲霉, 洛伐他汀, 根癌农杆菌, 遗传转化
Abstract: The high-yield lovastatin Monascus strain M120-1 screened by UV mutagenesis was used as the starting strain for extractioning total RNA. LovD gene was cloned by RT-PCR technique to construct expression vectors. The genetic transformation of Monascus spp. was obtained and the high-yield lovastatin transforming strains were screened by high performance liquid chromatography. The results showed that the total RNA of Monascus spp. was extracted and the expression vector was established, and twelve transformants were selected at random and 10 strains were successfully transformed by PCR. The lovastatin production was determined by HPLC and the yield of lovastatin was 0.55 mg/mL in one strain, which was 63% higher than that of the original strain. The strain was subcultured and had good genetic stability.
Key words: Monascus, lovastatin, Agrobacterium tumefaciens, genetic transformation
中图分类号:
Q78
沈小瑞， 王 钰， 陈达伟. 红曲霉LovD基因的克隆及转基因高产洛伐他汀菌株筛选[J]. 生物学杂志, doi: 10.3969/j.issn.2095-1736.2019.04.023.
SHEN Xiao-rui， WANG Yu， CHEN Da-wei. Cloning of monascus LovD gene and screening of transgenic high-yield lovastatin strains[J]. , doi: 10.3969/j.issn.2095-1736.2019.04.023.
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http://www.swxzz.com/CN/Y2019/V36/I4/23
