Optimization of the Sonication Assisted LiAc/SS-DNA/PEG Transformation of Saccharomyces cerevisiae

Main Article Content

Liu Beidong
Yang Qian*
Chen Dianfu
Zhou Qi
Song Jinzhu
Liu Heng

Abstract

The yeast and E. coli shuttle vector pYES2 and Saccharomyces cerevisiae H158 were used. The influence of the sonication treatment on yeast LiAc/SS-DNA/PEG transformation protocol was studied, and four main factors that influenced the transformation efficiency were optimized. Using the optimized protocol, the recombinant plasmid pYES2-CBHI was transform into H158. The results show that, sonication for 60s can increase the transformation efficiency to 1.1x103/µg plasmid. But when the treatment lasted more than 60s, the efficiency decreased sharply. Analysis of the orthogonal experiment indicate that group 5 (sonication time 60 seconds, incubation 40 minutes, SS-DNA 150µg, heat shock time 5 minutes) gave the best results. The main affecting factor was the quantity of the SS-DNA. The transformants of pYES2-CBHI were identified and the results indicate that the recombinant plasmid was transformed into the yeast cells with high efficiency.


Keywords: Saccharomyces cerevisiae, pYES2; transformation; sonication treatment


Corresponding author: E-mail: yangq@hope.hit.ecu.cn

Article Details

Section
Original Research Articles

References

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