真菌产人参皂苷葡萄糖苷酶基因的调取

Cloning of the ginsenoside glucosidase produced by fungus

根据已知的人参皂苷糖苷酶(GluGF)的N端氨基酸序列设计简并引物,从Absidia Sp.R84g菌株的总RNA出发,应用RACE技术,快速扩增测序得到cDNA3′末端与cDNA5′末端的未知序列,利用扩增测序所得到的cDNA5′末端和cDNA3′末端的序列设计引物,进行二次RT-PCR扩增并测序得到GluGF基因的全序列。回收PCR产物测序,得到GluGF基因的序列全长为2149 bp,其中5′UTR长度为82 bp,3′UTR长度为114 bp,GluGF的CDS区序列全长为1 923 bp,所编码的氨基酸序列的长度为640个氨基酸。

According to the N-terminal amino acid sequences of panaxsaponin-glucosidase produced by Absidia Sp.R84g,the complete sequence of GluGF was obtained using rapid-amplification of cDNA3′ and cDNA5′ ends,the RT-PCR.The length of complete sequence is 2 149 bp,3′ UTR is 114 bp 5′UTR is 82 bp,and the series of CDS length is 1 923 bp.