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小菜蛾颗粒体病毒PP31蛋白的原核表达及多克隆抗体制备

, PP. 129-133

Keywords: 小菜蛾颗粒体病毒,PP31,原核表达,多克隆抗体

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Abstract:

杆状病毒PP31是一种磷酸化的DNA结合蛋白。pp31基因存在于所有已完成测序的鳞翅目昆虫杆状病毒。用PCR方法从小菜蛾颗粒体病毒基因组扩增pp31基因,将其克隆至原核表达载体pET28a,转化大肠杆菌BL21(DE3)菌株。经IPTG诱导,在被转化菌株中表达产生出携带6×His标签的PP31蛋白。用ProBondTM树脂纯化的His-PP31融合蛋白大小约29kD左右。用该融合蛋白免疫新西兰大白兔,获得PlxyGVPP31蛋白抗血清。Westernblot分析显示,所获得的抗血清具有较强的特异性和高效价,可应用于PlxyGVPP31蛋白分析。

References

[1]  Hashimoto Y, Hayakawa T, Ueno Y, et al. Sequence analysis of the Plutella xylostella granulovirus genome[J]. Virology, 2000, 275 (2):358-372. [2] Guarino LA, Dong W, Xu B, et al. Baculovirus phosphoprotein pp31 is associated with virogenic stroma[J]. J Virol, 1992, 66(12): 7113-7120. [3] Guarino LA, Mistretta TA, Dong W. DNA binding activity of the baculovirus late expression factor PP31[J]. Virus Res, 2002, 90 (1-2):187-195. [4] Broussard DR, Guarino LA, Jarvis DL. Dynamic phosphorylation of Autographa californica nuclear polyhedrosis virus pp31[J]. J Virol, 1996, 70(10):6767-6774. [5] Lu A, Miller LK. The roles of eighteen baculovirus late expression factor genes in transcription and DNA replication[J]. J Virol, 1995, 69(2):975-982. [6] Rapp JC, Wilson JA, Miller LK. Nineteen baculovirus open reading frames, including LEF-12, support late gene expression[J]. J Virol, 1998, 72(12):10197-10206. [7] Yamagishi J, Burnett ED, Harwood SH, et al. The AcMNPV pp31 gene is not essential for productive AcMNPV replication or late gene transcription but appears to increase levels of most viral transcripts[J]. Virology, 2007, 365(1):34-47. [8] 黎路林, 刘攀峰, 王思民, 等. 小菜蛾颗粒体病毒PP31 与两种 宿主蛋白发生相互作用[J]. 病毒学报, 2012(1):15-22. [9] 张凌云, 卫静, 李长江, 等. 拟南芥转录因子NF-YC 的原核表 达及多克隆抗体制备[J]. 生物技术通报, 2012(3):57-62. [10] 史皆然, 郭晓雅, 师长宏, 等. 沙门菌外膜蛋白D 的原核表达 及多克隆抗体制备[J]. 生物技术通讯, 2010(4):497-500.

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