Cloning And Expression Of WZSP SLA-DQA In Escherichia Coli

Wuzhishan pig (WZSP) is special swine in china, and gene of WZSP is highly purification in the field of application, adapted to operation of genetic engineering; its heart, liver, Kinney, physiological and biochemical index of blood and organ size are similar to human, which is a kind of ideal animal model and heterogeneity organic transplant recipient of human.Major Histocompatibility Complex (MHC), also called SLA, MHC was divided into three types population:â… ,â…¡andâ…¢. Classical SLA-I( equal to human's HLA-A,-B,-C) included P1, P7, P14 etc, but the only P1 and P14 expressed protein product; classical SLA-â…¡mainly included DRA,DRB, DQA, DQB, DOB etc, but the only SLA-DR and SLA-DQ expressed protein; whileâ…¢located betweenâ… andâ…¡, at present,â…¢had 36 gene localizations.To clone and express WZSP SLA-DQA in Escherichia coli.To lay background for studying rejection mechanisms in xenotransplant.RT-PCR was performed to proliferate SLA-DQA genes, which were cloned into PGEM-T Easy vector for sequencing and analysis. The sequence of SLA-DQA cDNA showed no difference to the known sequence searched in the Gene bank, and then SLA-DQA cDNA was inserted into expression vector PET-28a(+). After enzyme digestion and sequence analysis, it was testified that the subclonal fragment inserted and the open reading frame all were correct, so the recombinant expression vector PET-28a(+)/SLA-DQA was successfully constructed. Then PET-28a(+)/SLA-DQA was induced by IPTG and target protein was identified by SDS-PAGE .The results indicated that the target protein expressed by Escherichia coli was coincident with expected.