|
|
|
| Screening of augmenter of liver regeneration-binding proteins by yeast-two hybrid technique |
| Jun Cheng, Lin Wang, Ke Li, Yin-Ying Lu, Gang Wang, Yan Liu, Yan-Wei Zhong, Hui-Juan Duan, Yuan Hong, Li Li, Ling-Xia Zhang and Ju-Mei Chen |
Beijing, China
From the Gene Therapy Research Center, Institute of Infectious Diseases, 302 Hospital of PLA, Beijing 100039, China (Cheng J, Wang L, Li K, Lu YY, Wang G, Liu Y, Zhong YW, Duan HJ, Hong Y, Li L, Zhang LX and Chen JM)
Correspondence: Jun Cheng, MD (Tel: 86-10-66933391; Fax: 86-10-63801283; Email: cj@genetherapy.com.cn; Website: www.genetherapy.com.cn) |
|
|
|
|
Abstract 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 2×YPDA medium. Diploid yeast was plated on a synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-α-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.
CONCLUSION: The successful cloning of gene of ALR interacting protein has paved the way for studying the physiological function of ALR and associated proteins.
|
|
|
|
|
|
|
|
|
2003, Vol. 2 
