山东医药2016，Vol.56Issue(19)：1-4,4.DOI:10.3969/j.issn.1002-266X.2016.19.001
酵母双杂交技术筛选宫颈癌 HeLa 细胞cDNA 文库中 FAM92 A1-289关联蛋白
Screening of interacting proteins of FAM92A1-289 in Hela cell cDNA library by using yeast two-hybrid technique
摘要
Abstract
Objective To screen the interacting proteins of FAM 92A1-289 in Hela cell cDNA library by yeast two-hy-brid technique.Methods The bait plasmid pGBKT7-FAM92A1-289 was constructed and then transformed into yeast com-petent cells AH109.Clontech GAL4 yeast two-hybrid assay was performed to screen the interacting proteins of FAM 92A1-289 in the Hela cell cDNA library .SD nutrient medium and X-a-Gal were used for selecting and screening .Moreover , back-cross was performed to confirm the interaction of identified proteins , and bioinformatics was used to analyze the se-quences of clones of positive β-galactosidase.Results The bait plasmid pGBKT7-FAM92A1-289 was successfully con-structed and expressed FAM92A1-289 in yeast cells, showing no toxicity and self-activation.Nine colonies were identified after β-galactosidase test and selection of SD/-Ade/-His/-Leu/-Trp plates containing X-a-Gal.Bioinformatics analysis shows four proteins had high repetition-rate, such as proliferating cell nuclear antigen ( PCNA), galectin-1, endoplasmic reticulum-golgi intermediate compartment marker 53 ( ERGIC-53 ) and recombinant human BCL 2-associated athanogene 1 (BAG1).Furthermore, back-cross proved there was interaction between the identified proteins .Conclusion The proteins interacting with FAM92A1-289 are successfully identified by yeast two-hybrid technique, which can provide clues for fur-ther investigation of the function of FAM 92A1-289.关键词
FAM92 A1-289/酵母双杂交技术/宫颈癌/HeLa细胞/增殖细胞核抗原/内质网高尔基体中间室标记物53/BCL2相关永生基因重组人BCL2相关永生基因1/半乳糖凝集素1Key words
FAM92A1-289/yeast two-hybrid technique/cervical carcinoma/HeLa cell/proliferating cell nuclear antigen/endoplasmic reticulum-golgi intermediate compartment marker 53/recombinant human BCL2-associated athanogene 1/galectin-1分类
生物科学引用本文复制引用
沈君豪,方娟,郭兴荣,桂卉,涂汉军,阮绪芝..酵母双杂交技术筛选宫颈癌 HeLa 细胞cDNA 文库中 FAM92 A1-289关联蛋白[J].山东医药,2016,56(19):1-4,4.基金项目
湖北省自然科学基金资助项目（2012FFB02001）。 （）
