[1]杨琳,耿明星,江乾森,等.家蚕天然Hemolin蛋白原核表达[J].浙江理工大学学报,2020,43-44(自科五):664-670.
YANG Lin,GENG Mingxing,JIANG Qiansen,et al.Prokaryotic expression of natural Hemolin protein from Bombyx mori[J].Journal of Zhejiang Sci-Tech University,2020,43-44(自科五):664-670.
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家蚕天然Hemolin蛋白原核表达(
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YANG Lin,GENG Mingxing,JIANG Qiansen,et al.Prokaryotic expression of natural Hemolin protein from Bombyx mori[J].Journal of Zhejiang Sci-Tech University,2020,43-44(自科五):664-670.
)浙江理工大学学报[ISSN:1673-3851/CN:33-1338/TS]
- 卷:
- 第43-44卷
- 期数:
- 2020年自科五期
- 页码:
- 664-670
- 栏目:
- 出版日期:
- 2020-09-18
文章信息/Info
- Title:
- Prokaryotic expression of natural Hemolin protein from Bombyx mori
- 文章编号:
- 1673-3851 (2020) 05-0664-07
- 分类号:
- Q71
- 文献标志码:
- A
- 摘要:
- Hemolin蛋白是一种类免疫球蛋白，属于昆虫的一种重要的先天性免疫球蛋白，家蚕（Bombyx mori）Hemolin蛋白与其他昆虫的序列具有高度的同源性，为了进一步分析家蚕Hemolin蛋白的功能，以杆状病毒感染的家蚕蚕蛹为原料，通过抗体亲和层析获得高纯度的天然Hemolin蛋白。结果表明：重组表达菌株pET32ahemolin经终浓度为01 mmol/L IPTG低温诱导后可在大肠杆菌BL21中成功表达；通过镍离子金属螯合亲和层析并经200 mmol/L咪唑洗脱可以获得较纯的重组Hemolin蛋白；免疫新西兰大白兔产生特异性多克隆抗体通过蛋白A亲和层析柱纯化后可获得高纯度的Hemolin抗体，并经抗体亲和层析获得的天然Hemolin蛋白纯度高，可用于对Hemolin蛋白功能的进一步研究。文章提供的分离纯化高纯度家蚕天然Hemolin蛋白的方法能够推进天然免疫蛋白的纯化，为进一步研究家蚕天然Hemolin蛋白的晶体结构、蛋白质抑菌功能以及抗病毒活性奠定基础。
参考文献/References:
［1］ Wang Q, Ren M, Liu X, et al. Peptidoglycan recognition proteins in insect immunity［J］. Mol Immunol, 2019, 106(45): 69-76.
［2］ Ferrandon D, Imler J L, Hetru C, et al. The Drosophila systemic immune response: Sensing and signalling during bacterial and fungal infections［J］. Nat Rev Immunol, 2007, 7(11): 862-874.
［3］ Levitin A, Whiteway M. Drosophila innate immunity and response to fungal infections［J］. Cell Microbiol, 2008, 10(5): 1021-1026.
［4］ Su X D, Gastinel L N, Vaughn D E, et al. Crystal structure of hemolin: A horseshoe shape with implications for homophilic adhesion［J］. Science, 1998, 281(5379): 991-995.
［5］ Qian C, Wang F, Zhu B, et al. Identification of a hemolin protein from actias selene mediates immune response to pathogens［J］. Int Immunopharmacol, 2017, 42(1): 74-80.
［6］ Sun Y, Wang L, Qian C, et al. Molecular cloning and expression analysis of a hemolinlike molecule from Antheraea pernyi［J］. Int Immunopharmacol, 2015, 26(1): 65-71.
［7］ Schmidt O, Soderhall K, Theopold U, et al. Role of adhesion in arthropod immune recognition［J］. Annu Rev Entomol, 2010, 55(55): 485-504.
［8］ Li W, Terenius O, Hirai M, et al. Cloning, expression and phylogenetic analysis of hemolin, from the chinese oak silkmoth, Antheraea pernyi［J］. Dev Comp Immunol, 2005, 29(10): 853-864.
［9］ Yu X Q, Kanost M R. Developmental expression of Manduca sexta hemolin［J］. Arch Insect Biochem, 1999, 42(3): 198-212.
［10］ Terenius O. HemolinA lepidopteran antiviral defense factor?［J］. Dev Comp Immunol, 2008, 32(4): 311-316
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备注/Memo
- 备注/Memo:
-
收稿日期：2020-06-08
网络出版日期： 2020-07-09
基金项目：浙江省重点研发计划项目(2019C02043)
作者简介：杨琳(1994-)，女，江苏盐城人，硕士研究生，主要从事动物疫病防控研究
通信作者：舒建洪 E-mail: shujianhong@zstu.edu.cn
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