Screening of augmenter of liver regeneration-binding proteins by yeast-two hybrid technique.
OBJECTIVE
To investigate the biological function of augmenter of liver regeneration (ALR), we used yeast-two hybrid technique to detect proteins in hepatocytes interacting with ALR.


METHODS
ALR bait plasmid was constructed by using yeast-two hybrid system 3, then transformed into yeast AH109. The transformed yeast was mated with yeast Y187 containing liver cDNA library plasmid in a 2XYPDA medium. Diploid yeast was plated on a synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selection and screening. After extracting and sequencing of the plasmid from blue colonies. Analysis was performed by bioinformatics.


RESULTS
Of 36 colonies sequenced, 14 are metallothionein, 12 albumin, and 3 selenoprotein P. One colony is a new gene with unknown function.


CONCLUSIONS
The successful cloning of gene of ALR interacting protein has paved the way for studying the physiological function of ALR and associated proteins.
Cheng
Jun
J
Gene Therapy Research Center, Institute of Infectious Diseases, 302 Hospital of PLA, Beijing 100039, China. cj@genetherapy.com.cn
Wang
Lin
L
Li
Ke
K
Lu
Yin-Ying
YY
Wang
Gang
G
Liu
Yan
Y
Zhong
Yan-Wei
YW
Duan
Hui-Juan
HJ
Hong
Yuan
Y
Li
Li
L
Zhang
Ling-Xia
LX
Chen
Ju-Mei
JM
D000818:Animals; D001483:Base Sequence / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics; D019976:Cloning, Organism / Q000379:methods; D015723:Gene Library; D022781:Hepatocytes / Q000502:physiology*; D006824:Hybridization, Genetic / Q000502:physiology; D008115:Liver Regeneration / Q000235:genetics* / Q000502:physiology; D008403:Mass Screening / Q000379:methods*; D011506:Proteins / Q000235:genetics*; D015003:Yeasts; D050862:Oxidoreductases Acting on Sulfur Group Donors
D002352:Carrier Proteins; D011506:Proteins; C088825:GFER protein, rat; D050862:Oxidoreductases Acting on Sulfur Group Donors