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High level expression of glucose-6-phosphate dehydrogenase gene <Emphasis Type="Italic">PsG6PDH</Emphasis> from <Emphasis Type="Italic">Populus suaveolens</Emphasis> in <Emphasis Type="Italic">E. coli</Emphasis>
Authors:Lin?Yuan-zhen  Email author" target="_blank">Zhang?Zhi-yi?Email author  Lin?Shan-zhi  Zhang?Qian  Wang?Xin
Institution:(1) Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry University, 100083 Beijing, P. R. China
Abstract:In order to investigate the functions of the gene PsG6PDH and the mechanisms underlying freezing tolerance of Populus suaveolens, the recombinant expression vector pET-G (pET30a-G6PDH), which contained full encoding region of PsG6PDH gene, was established. The recombinant was identified by lawn-PCR and double enzyme digestion and then transformed into expression host XA90 and induced by isopropyl-a-D-thiogalactoside (IPTG) to express 100 kD polypeptide of G6PDH fusion protein. The results showed that the expressed amount of the fusion protein culminated after 1 mmol·L−1 IPTG treatment for 4 h and that pET-G product was predominately soluble and not extra-cellular secreting. Supported by the National Natural Science Foundation of China (Grant No. 30271093)]
Keywords:Populus suaveolens  glucose 6-phosphate dehydrogenase  PsG6PDH  prokaryotic expression
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