The effective expression of xylanase gene in Candida utilis by 18S rDNA targeted homologous recombination in pGLR9K.
In order to test whether 18S rDNA can influence positively xylanase gene effective expression in the yeast of Candida utilis, a targeting vector pGLR9K-XA was constructed by adding an interested gene xynA from Streptomyces olivaceoviridis into the vector pGLR9K which is constructed by ourselves. pGLR9K contains the 18S rDNA, GAP promoter and CYH resistance gene sequence, all of which is from C. utilis. Then the vector pGLR9K-XA was transformed into C. utilis. To test the vector and transformed system, PCR, Southern blot and DNS methods were used. The results showed that xylanase gene can be detected in the chromosome DNA of recombinant C. utilis and the enzyme activity of xylanase is up to 60 IU ml(-1) in the study. It is suggested that this system can be used to express exogenous genes in C. utilis as a bioreactors. This is the first report that xylanase gene was expressed in C. utilis.
Wei
Wang
W
Insititute of Microbiology of Xinjiang Academy of Agricultural Sciences, 830091, Urumqi, China. whangwei0718@126.com
Hong-Lan
Yang
Y
HuiFang
Bao
B
Daoyuan
Zhang
Z
Qi-mu-ge
Shan
S
Woof
Andrew J
AJ
D002175:Candida / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D043364:Endo-1,4-beta Xylanases / Q000096:biosynthesis* / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D059765:Homologous Recombination / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012337:RNA, Ribosomal, 18S / Q000235:genetics*
D012337:RNA, Ribosomal, 18S; D043364:Endo-1,4-beta Xylanases