[乙型肝炎病毒e抗原相互作用蛋白基因的筛选]

[Screening of the genes of hepatitis B virus e antigen interacting proteins].

作者信息

Li Bo-an, Shu Cui-li, Hou Jun, Qi Yang, Li Jing, He Wei-ping, Cheng Yun

机构信息

Department of Immunology, Institute of Infectious Disease of The No. 302 Hospital, Beijing 100039, China.

出版信息

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2005 Sep;19(3):271-4.

PMID:16261214

Abstract

OBJECTIVE

To screen and clone the genes in hepatocytes which encode protein that can interact with hepatitis B e antigen(HBeAg) by yeast-two hybridization.

METHODS

Recombined HBeAg bait plasmid (pGBKT7-eAg) was transformed into yeast AH l09, followed by mating with yeast Yl87 containing liver cDNA library plasmid in 2 x YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-Ade-His) which contains X-a-gal for selecting positive blue clones. Then positive clones were selected and plasmids were prepared and sequenced. Finally, bioinformatics analysis was performed.

RESULTS

Totally 245 positive colonies were selected and 101 colonies were sequenced. Through sequences alignment, 6 novel genes and 35 recorded genes were screened.

CONCLUSION

Genes of HBeAg interacting proteins have been cloned successfully, which brings some new clues for further studies on the biological functions of HBeAg and the related proteins.

摘要

目的

通过酵母双杂交技术筛选并克隆肝细胞中编码能与乙型肝炎e抗原（HBeAg）相互作用蛋白的基因。

方法

将重组的HBeAg诱饵质粒（pGBKT7-eAg）转化至酵母AH109，随后在2×YPDA培养基中与含有肝脏cDNA文库质粒的酵母Y187进行交配。将二倍体酵母接种于含有X-α-半乳糖的合成缺陷营养培养基（SD/-Trp-Leu-Ade-His）上，以筛选阳性蓝色克隆。然后挑选阳性克隆，制备质粒并进行测序。最后进行生物信息学分析。