酿酒酵母BJ3505 NHX1基因突变株的构建及功能验证

doi:10.13560/j.cnki.biotech.bull.1985.2018-0561

摘要/Abstract

摘要： 构建酿酒酵母NHX1基因单缺失突变株,验证其在酿酒酵母BJ3505抵抗逆境及液胞融合中的功能。利用同源重组技术构建BJ3505 NHX1基因的插入失活突变株BJ3505Δnhx1,并构建互补菌株BJ3505Δnhx1-C,对酿酒酵母NHX1基因在逆境胁迫及液胞融合中的功能进行验证研究。结果表明：与互补菌株BJ3505Δnhx1-C和野生型相比,突变株BJ3505Δnhx1 抗逆能力减弱,液胞融合受影响,导致多液胞存在。利用同源重组法成功的构建了NHX1基因缺失突变株;酿酒酵母NHX1作为重要的离子反向转运体,在酿酒酵母BJ3505抵抗外界胁迫环境和液胞融合过程中发挥着重要的作用。

关键词: 酿酒酵母, NHX1, 逆境胁迫, 液胞融合

Abstract: A Saccharomyces cerevisiae mutant with deficient NHX1 gene was obtained for verifying its function in S. cerevisiae BJ3505's resistance to stress and vacuole fusion. The BJ3505Δnhx1,the mutant of BJ3505 with inactivated NHX1,was cloned via homologous recombination. Then the complementary strain BJ3505Δnhx1-C was constructed to validate the function of NHX1 in stress tolerance and vacuole fusion. As results,mutant BJ3505Δnhx1 had a much lower ability in stress tolerance and an increase in cells harboring multiple vacuoles was observed,compared with the BJ3505Δnhx1-C and the wild one. Conclusively,the NHX1gene-deleted mutant strain was constructed successfully by homologous recombination. As a crucial ion antiporter,NHX1 plays an important role in the stress resistance and vacuole fusion of S. cerevisiae BJ3505.

Key words: Saccharomyces cerevisiae, NHX1, adversity stress, vacuole fusion

王立光, 陈军, 叶春雷, 罗俊杰. 酿酒酵母BJ3505 NHX1基因突变株的构建及功能验证[J]. 生物技术通报, 2018, 34(12): 152-158.

WANG Li-guang, CHEN Jun, YE Chun-lei, LUO Jun-jie. Mutant Construction and Functional Validation of NHX1 in Saccharomyces cerevisiae BJ3505[J]. Biotechnology Bulletin, 2018, 34(12): 152-158.

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