[Cloning and Expression of Various Deletants of Gsalpha Gene in Escherichia Coli]

Zhongguo Shi Yan Xue Ye Xue Za Zhi 2000 Dec;8(4):251-254

Hematological Department, The General Hospital of Beijing Military Command, Beijing 100700, China; Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100850, China; Institute of Molecular Biology, General Hospital of PLA, Beijing 100853, China.

Gsalpha gene mutation has been discovered in some human tumors. In our previous studies, three novel deletants of Gsalpha gene, Gsalpha L-1(500 bp), Gsalpha L-2(300 bp), and Gsalpha L-3(200 bp), and wild type Gsalpha-4(1 200 bp) were found in human leukemia cell lines and detected in leukemic cells from patients with acute leukemia. To investigate the construction, function and biological significance of the deletants, the plasmids of Gsalpha L-1, Gsalpha L-2 and wild Gsalpha-4 were transformed into E. coli DH5, amplified by PCR, and cloned in expression vector pET22b(+), and then transformed into E. coli, respectively. As a result, higher levels of expression of three recombinants were obtained in form of inclusion bodies. The results suggested that these Gsalpha isoforms have an open reading frame of gene and can be expressed in vitro. The data lay a foundation to study the relation of Gsalpha gene to leukemogenesis.

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December 2000
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