title: Cloning and expression of the gene of augmenter of liver regeneration in yeast cells.
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 weight was identical to the theoretical value of 15 000 Da; the protein was found inside the yeast cells.
  
  
  CONCLUSIONS
  The successful expression of ALR in yeast cells makes it possible to study further on its biological function.
authors: Gene Therapy Research Center, Institute of Infectious Diseases, 302 Hospital of PLA, Beijing 100039, China. cj@genetherapy.com.cn
authors: Jun
authors: J
authors: Cheng
authors: Lin
authors: L
authors: Wang
authors: Ke
authors: K
authors: Li
authors: Yin-Ying
authors: YY
authors: Lu
authors: Yan
authors: Y
authors: Liu
authors: Hui-Juan
authors: HJ
authors: Duan
authors: Yuan
authors: Y
authors: Hong
authors: Gang
authors: G
authors: Wang
authors: Li
authors: L
authors: Li
authors: Ling-Xia
authors: LX
authors: Zhang
mesh_terms: D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D018076:DNA, Complementary; D015870:Gene Expression; D006801:Humans; D008113:Liver Neoplasms; D008115:Liver Regeneration / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D011506:Proteins / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D015183:Restriction Mapping; D015003:Yeasts; D050862:Oxidoreductases Acting on Sulfur Group Donors
chemical_list: D018076:DNA, Complementary; D011506:Proteins; D011994:Recombinant Proteins; C088825:GFER protein, rat; D050862:Oxidoreductases Acting on Sulfur Group Donors