酵母菌生长曲线的测定及其转葡萄芪合酶基因重组菌遗传稳定性的检测

Measurement of Yeast Growth Curve and Genetic Stability Examination of Recombinant Yeast Transferred Grape Stilbene Synthase(STS)

采用分光光度计比浊法测定酿酒酵母INVScl的生长曲线，确定菌体生长规律；并以此为依据，制备酿酒酵母感受态细胞，进行葡萄芪合酶基因酿酒酵母重组质粒的转化；采用菌落PCR法快速鉴定阳性重组子，并进一步检测重组质粒在重组菌中的稳定性。结果表明，于酵母菌对数生长中期（OD600为0．8时）进行重组质粒的转化，可以得到较高的转化效率；在0～72h发酵培养过程中重组质粒在宿主菌中的稳定性达到100％。这为后续工作开展葡萄芪合酶基因在酿酒酵母中的诱导表达研究奠定了技术基础，

Yeast growing rule was confirmed by measuring the growth curve of INVScl by spectro-photometer turbidity. According to this growing curve, yeast competent cells were made up and were transformed by the yeast recombinant plasmid containing stilbene synthase gene derived from grape. The positive recombinant vector was rapidly identified by using colony PCR. Then, the stability of recombinant vector in recombinant yeast was examined, The resuits stated, at the mid-logarithmic phase （ OD600 of 0.8） the transformation of recombinant plasmid was high-efficient, and during 0 - 72 h cultivation pefled the stability rate of the recombinant plasmid in yeast reached 100%. It may lay the technical foundation for the further inducible expression of SIS gene in S. cerevisiae.