Cloning of the LAC4 Promoter from K. lactis and Studies on the Function of its UASII.
The -661 - +21 bp region of the LAC4 gene form K. lactis CBS141 was obtained by PCR technique and fused with the lacZ gene from E. coli to construct and expression vector YFD 114. It was then transformed into K. lactis Y167. The function of the cloned LAC4 promoter was tested by induction with galactose, lactose, sorbitol or IPTG. The results revealed that galactose had higher inducing effect than lactose while sorbitol and IPTG had no inducing effect. The insertion of chemically synthesized LAC4 UASII at just the upstream of the cloned LAC4 promoter could increase its basic and induced expression level. The extents of increase were different with the orientation and the copy number of the inserted UASII.
Bao
Wei-Guo
WG
Institute of Genetics, Fudan University, Shanghai 200433, China.
Chen
Yu
Y
Huo
Ke-Ke
KK
Li
Yu-Yang
YY