Culture of Escherichia coli in SOC medium improves the cloning efficiency of toxic protein genes.
In this paper, we report a useful protocol for cloning toxic protein genes. Use of the SOC medium, which is a glucose-containing rich medium, significantly improved the transformation efficiency of a recombinant plasmid containing a toxic plant subtilase SaSBT1 cDNA. Both glucose and rich nutrients present in the SOC medium prevented the unintended activation of the lac promoter carried on the cloning vector, and led to significantly improved transformation efficiency of recombinant plasmids containing toxic protein genes and an increased rate of transformant growth.
Sun
Qiao-Yang
QY
State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, Guangdong, China.
Ding
Ling-Wen
LW
He
Liang-Liang
LL
Sun
Yong-Bin
YB
Shao
Jun-Li
JL
Luo
Ming
M
Xu
Zeng-Fu
ZF
D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular / Q000187:drug effects / Q000379:methods; D003470:Culture Media / Q000737:chemistry* / Q000494:pharmacology*; D004926:Escherichia coli / Q000166:cytology / Q000187:drug effects* / Q000235:genetics*; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D050296:Microbial Viability / Q000187:drug effects*; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D032322:Solanum / Q000235:genetics; D014169:Transformation, Bacterial / Q000187:drug effects
D003470:Culture Media; D010940:Plant Proteins; D011994:Recombinant Proteins; D005947:Glucose