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| Cloning and expression of the gene of augmenter of liver regeneration in yeast cells |
| Jun Cheng, Lin Wang, Ke Li, Yin-Ying Lu, Yan Liu, HuI-Juan Duan, Yuan Hong, Gang Wang, Li LI and Ling-Xia Zhang |
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, Liu Y, Duan HJ, Hong Y, Wang G, Li L and Zhang LX)
Correspondence: Jun Cheng, MD (Tel: 86-10-66933391; Fax: 86-10-63801283; Email: cj@genetherapy.com.cn) |
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Abstract Objective: To study the function of augmenter of liver regeneration (ALR) as a regulatory factor that specifically stimulates hepatic cell regeneration, we constructed yeast expressive vector of ALR and expressed it in yeast cells.
Methods: Total RNA was extracted from HepG2 cells, and reverse transcription polymerase chain reaction (RT-PCR) was performed to amplify the coding region of ALR. The products were cloned into PGEM-T vector and sequenced, then cloned into PGBK T7 vector. The recombinant plasmid PGBK T7-ALR was transformed into yeast AH109. The yeast protein was extracted and analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting hybridization technique.
Results: DNA sequencing results confirmed that the coding region of ALR was correctly inserted into the yeast expression vector, and Western blotting assay showed that recombinant ALR was successfully expressed in yeast. Its molecular weightwas identical to the theore tical value of 15 000 Da; the pro tein was found inside the yeast cells.
Conclusion: The successful expression of ALR in yeast cells makes it possible to study further on its Biological function.
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2002, Vol. 1 
