Screening of Proteins Binding to HBsAg from Human Pancreas cDNA Library by Yeast Two-hybrid System / 环境与健康杂志

ABSTRACT

Objective To screen proteins in pancreas cDNA library interacting with HBsAg protein.Methods After being amplified and identified,the human pancreas cDNA library plasmid was transformed into yeast Y187.Thereafter,the bait plasmid(PGBKT7-HBsAg) was transformed into yeast cells AH109.Then,two different transformed yeast cells were mated.The diploid yeast cells were plated on synthetic dropout nutrient medium(SD/-Trp/-Leu/-His/-Ade) and that medium contained X-?-gal for selection two times.After plasmids of true positive blue colonies were extracted and transformed to E.coli,the results were analyzed by DNA sequencing and blast searching in GenBank database.Results Six proteins binding to HBsAg were screened,including aconitase(ACO1),CDK5RAP3,carboxypeptidase,etc.Conclusion Most of these proteins play significant roles in the evolution of 2 type diabetes mellitus,which may supply some new clues for exploring the pathogenesis of abnormality of lipid metabolism and tglycometabolism in the patients with chronic hepatitis virus infection.