严永敏, 许文荣, 朱海波, 张岭, 费仁仁, 王晓良, 钱晖, 郝顺祖, 王文兵. 人磷酸二酯酶同工酶3A在昆虫杆状病毒系统中的表达J. 药学学报, 2005, 40(9): 810-813.
引用本文: 严永敏, 许文荣, 朱海波, 张岭, 费仁仁, 王晓良, 钱晖, 郝顺祖, 王文兵. 人磷酸二酯酶同工酶3A在昆虫杆状病毒系统中的表达J. 药学学报, 2005, 40(9): 810-813.
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YAN Yong-min, XU Wen-rong, ZHU Hai-bo, ZHANG Ling, FEI Ren-ren, WANG Xiao-liang, QIAN Hui, HAO Shun-zu, WANG Wen-bing. Expression of human phosphodiesterase 3A gene using baculovirus expression system in insect cellJ. Acta Pharmaceutica Sinica, 2005, 40(9): 810-813.
Citation: YAN Yong-min, XU Wen-rong, ZHU Hai-bo, ZHANG Ling, FEI Ren-ren, WANG Xiao-liang, QIAN Hui, HAO Shun-zu, WANG Wen-bing. Expression of human phosphodiesterase 3A gene using baculovirus expression system in insect cellJ. Acta Pharmaceutica Sinica, 2005, 40(9): 810-813.
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人磷酸二酯酶同工酶3A在昆虫杆状病毒系统中的表达

Expression of human phosphodiesterase 3A gene using baculovirus expression system in insect cell

    摘要
  • 摘要: 目的利用杆状病毒表达系统探讨人磷酸二酯酶同工酶3A(HPDE3A)在Tn细胞中的表达。方法用重组HPDE3A杆状病毒感染昆虫Tn细胞进行蛋白表达，通过RT-PCR，SDS-PAGE和Western blotting技术检测HPDE3A表达情况。结果重组杆状病毒在感染Tn细胞后形态变化明显，获得表达产物,RT-PCR扩增结果证实了Tn细胞中HPDE3A的mRNA水平增高，Tn细胞能够表达出与HPDE3A多克隆抗体结合的蛋白，蛋白分子质量约为120 kD。结论HPDE3A在昆虫杆状病毒表达系统中可以稳定表达，为进一步研究HPDE3A的生物学活性及筛选HPDE3A抑制剂奠定了基础。

     

    Abstract: AimTo investigate the expression of recombinant human phosphodiesterase 3A (HPDE3A) using baculovirus expression system in Tn cell line. MethodsThe HPDE3A cDNA was recombined with baculovirus, and then the recombinant was transfected into Tn cell line. The expression of HPDE3A in Tn cell line was detected and identified by the RT-PCR, SDS-PAGE and Western blotting. ResultsThe recombinant HPDE3A protein was stably expressed in Tn cell line and detected by the distinct morphological changes of Tn cell, RT-PCR, SDS-PAGE and Western blotting using polyclonal antibody. The MW of the recombinant protein was about 120 kD. ConclusionRecombinant HPDE3A can be expressed in Tn cell line using the baculovirus expression system, and thus provided the basic material for studying its bioactivity and application in screening for HPDE3A inhibitor.

     

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