酵母克隆基因的转化和稳定性检测

采用放射性探针分子杂交技术鉴定及选出重组DNA的可靠转化子。实验结果表明:双倍体受体的转化效率约为单倍体的2倍;含有酵母染色体端粒区GT重复顺序的重组质粒的转化效率约为含染色体内部区GT重复顺序重组质粒的3倍;在克隆基因的稳定性方面,整合型转化子要比自主复制型转化子高得多。因此,用酿酒酵母作基因工程生产菌时以双倍体为好。经分析并证实,含酵母染色体端粒区GT重复顺序的重组子有些具有多靶整合作用,这种整合转化在基因工程中有实用价值。

Transformation of Cloned Gene in Yeast and Detection of Stability of Transformants

In this paper the application of Liac transfomation method to intact yeast cells was reported. Transformants of recombinant DNA was confirmed by molecular hybridization technique Results showed that the transformation efficiency of diploid was about twice that of haploid, the transformation efficiency of telomeric relion (GT) n-containing plasmids and the stability of integrative transformants was much higher than that of autonomous replicating transformants. The above experimental results suggest that the diploid yeast is better for gene cloning production and that the telomeric region(GT) n-containing plasmids are capsble of integrating to multiple sites on chromosomes. Multitarget integration is very useful in genetic egineering.