A novel aberrant splicing of G(s alpha) transcript in human leukemia cell lines

作者：Ye QN^*; Yao XA; Wang HL; Zhang S; Liu HT; Su GF; Hang CF; Zhou TC

来源：Chinese Science Bulletin, 1999, 44(4): 340-344.

DOI：10.1007/BF02885487

• G(s alpha)
• splicing
• leukemia
• cloning and expression

The alpha subunit of the stimulatory G protein, G(s alpha), is involved in the stimulation of adenylate cyclase pathway of signal transduction. The various G(s alpha) mRNAs are generated either by alternative splicing or by using alternative promoter. A novel aberrant splicing of G(s alpha) transcript in human leukemia cell lines is reported. The entire coding region of G(s alpha) gene obtained, referred to as G(s alpha Leu) was amplified by PCR with cDNA from Jurkat human leukemia cDNA library or the reverse-transcribed first strand cDNA from human leukemia cell line HL-60 as a template. The result of DNA sequencing indicated that, compared with originally reported G(s alpha), G(soLeu) has two in-frame deletions, one in amino acid residues 23-249 and the other in amino acid residues 254-260, suggesting that G(s alpha Leu) may play an important role in leukemia. The complete coding region of G(s alpha Leu) was inserted downstream of the thrombin site of pGEX-2T fusion protein expression vector and expressed in the form of GST/Gs(alpha Leu) at high level, which was then purified to be electrophoretically pure by one-step affinity chromatography. The expression of G(s alpha Leu) lays foundation for the study of its function.

• 出版日期1999-2