[Cloning and expression of maltooligosyltrehalose synthase from Sulfolobus acidocaldarius in Escherichia coli].
The gene of MTSase (maltooligosyltrehalose synthase) from Sulfolobus acidocaldarius ATCC49426 was amplified by PCR. The primers were designed according to the published sequence of homologous gene from Sulfolobus acidocaldarius ATCC33909. This gene was inserted into the plasmid pBV220 and the resultant recombinant plasmid pBV220-GT was transformed to E. coli DH5 alpha. The activity of recombinant enzyme was about 10 u/g(wet cell). In order to improve the expression level of target protein, some nucleotides in the 3' and 5' of the gene were modified to optimize the second structure of mRNA by PCR amplification using the new primers devised according to the biosoftware GOLDKEY2.0. As a result, the activity of recombinant enzyme increase to 19.8 u/g(wet cell). Then, the helping plasmid pUBS520 which carried the gene encoding the tRNA of rare codons AGG and AGA was transformed to the recombinant strain. But it took little effect.
Wang
H
H
Institute of Microbiology, Chinese Academy of Science, Beijing 100080, China. wujinw5@sina.com.cn
Chen
W
W
Wu
J
J
Jin
C
C
D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D016945:Sulfolobus acidocaldarius / Q000201:enzymology*
D005964:Glucosyltransferases; C099084:maltooligosyl trehalose synthase