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Minimal Set of Metabolic Pathways Suggested from the Genome of Onion Yellows Phytoplasma 总被引:1,自引:0,他引:1
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AIM and METHODS: Total RNA was extracted from 6th rat subcultured pulmonary artery smooth muscle cells(PASMC) exposed to continual chronic hypoxia or normoxia and the effects of chronic hypoxia on the changes of Kv1.3,Kv2.1,Kv3.1 mRNA in cultured PASMC induced by acute hypoxia were studied by semiquantitative RT-PCR in vitro. RESULTS:①Kv1.3,Kv2.1,Kv3.1 genes were found to be expressed in PASMC of rats exposed either to hypoxia or normxia.②The expression of Kv2.1 and Kv3.1 in 6th subcultured of PASMC in normaxia group could be upregulated by exposure to acute hypoxia,the levels of Kv2.1 and Kv3.1 mRNA were significantly increased from 0.646±0.092, 0.782±0.104 to 1.059±0.134, 0.985±0.116,respectively (P<0.01,n=5). ③PASMC cultured continuously in chronic hypoxia for 6 subcultures and then exposed to normoxia for 12 h,thereafter the expression of Kv2.1 and Kv3.1 were downregulated by acute hypoxia for 6 hours.The level of Kv2.1 mRNA was significantly decreased from 1.008±0.117 to 0.649±0.097 (P<0.01,n=5). CONCLUSION:Kv2.1,Kv3.1 genes might be oxygen sensitive genes.Chronic hypoxia might change the response of these Kv genes of PASMC to acute hypoxia and down-regulate its expression,which might probably decrease the role of Kv in HPV. 相似文献
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AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors. 相似文献
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肥城桃组培苗诱导、基因转化及其增殖 总被引:11,自引:0,他引:11
红里、白里是肥城桃的两个优良品种,但不耐贮藏。为试图利用基因工程解决这一问题,先通过组培将其胚、胚乳、子叶的愈伤组织分别诱导再生植株和芽剥离出茎尖培育成苗。再将肥城桃反义PG基因,通过农杆菌介导转化组培苗,转化苗在含卡那霉素培养基上筛选,获得抗卡那霉素的转基因苗。用研制的增殖培养基培养4~7周,一个苗可生长出14~18分枝的苗,解决了转基因苗获得率低的问题,也满足了对转基因株系进行检测和从组培室到温棚到田间果园过渡栽培和芽接的需求,提高了繁育速率。 相似文献