Screening of interacting proteins of FAM92A1-289 in Hela cell cDNA library by using yeast two-hybrid technique
SHEN Junhao
FANG Juan
GUO Xingrong
GUI Hui
TU Hanjun
RUAN Xuzhi
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.
Keywords：FAM92A1-289yeast two-hybrid techniquecervical carcinomaHeLa cellproliferating cell nuclear antigenendoplasmic reticulum-golgi intermediate compartment marker 53recombinant human BCL2-associated athanogene 1galectin-1
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 1-4 )