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中国农学通报

中国农学通报 ›› 2015, Vol. 31 ›› Issue (23): 30-35.doi: 10.11924/j.issn.1000-6850.casb15010148

所属专题： 生物技术； 水产渔业

• 水产 渔业 • 上一篇    下一篇

对虾白斑综合症病毒(WSSV)蛋白激酶wsv083的初步研究

秦崇涛,杨丰   

  1. 福建中医药大学中西医结合学院,国家海洋局第三海洋研究所海洋生物遗传资源重点实验室
  • 收稿日期:2015-01-21 修回日期:2015-06-23 接受日期:2015-06-25 出版日期:2015-08-20 发布日期:2015-08-20
  • 通讯作者: 杨丰
  • 基金资助:
    国家自然科学基金(31072243)

Study on Protein Kinase wsv083 of White Spot Syndrome Virus

  • Received:2015-01-21 Revised:2015-06-23 Accepted:2015-06-25 Online:2015-08-20 Published:2015-08-20

PDF (PC)

396

摘要： 为探讨VP19和VP28是否由蛋白激酶wsv083催化发生磷酸化，将wsv083、VP19和VP28分别进行原核表达。将wsv083蛋白、wsv083表达菌株的裂解液上清以及去磷酸化的wsv083蛋白分别和2种膜蛋白相互作用，以及将wsv083、VP19和VP28的表达质粒共转化至同一菌株中进行表达。结果均未能发现VP19和VP28被磷酸化，但发现wsv083蛋白自身具有苏氨酸和酪氨酸磷酸化的现象。改变wsv083表达质粒，wsv083蛋白同样出现了磷酸化现象。而在将wsv083的序列改变使其失去激酶活性后，表达出的wsv083蛋白不再具有磷酸化的现象。因此认为VP19和VP28的磷酸化不是由wsv083催化产生，wsv083能够自我磷酸化。

关键词: 油茶饼粕, 油茶饼粕, 提取工艺, 优化

Abstract: To study the phosphorylation catalyze effect of wsv083, wsv083, VP19 and VP28 were sub-cloned into a pET28a vector and expressed in the prokaryotic expression system. The obtained protein wsv083, supernatant of wsv083 expression strain lysate and dephosphorylation wsv083 were used to interact with VP19 and VP28. Plasmids of wsv083, VP19 and VP28 were transformed into the same host strain. The results showed that phosphorlation of VP19 and VP28 were not found, while wsv083 itself displayed threonine and tyrosine phosphorylation. The author changed expression plasmid of wsv083, but wsv083 still presented the same phosphorylation. Furthermore, when the sequence of wsv083 was mutated to make its kinase activity lost, the expressed wsv083 did not have any phosphorylation. In conclusion, the results presented that phosphorylation of VP19 and VP28 were not catalyzed by wsv083, but wsv083 was able to make self-phosphorylation.