大劣按蚊卵黄蛋白原基因cDNA全长序列扩增及功能域蛋白的表达与纯化研究

于莎莎; 胡辉; 秦杰; 王静; 刘婷; 刘婷婷; 王英

doi:10.16016/j.1000-5404.202012233

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大劣按蚊卵黄蛋白原基因cDNA全长序列扩增及功能域蛋白的表达与纯化研究

基础医学 | 更新时间：2026-01-09

- 大劣按蚊卵黄蛋白原基因cDNA全长序列扩增及功能域蛋白的表达与纯化研究
- Amplification of full-length cDNA sequence of Anopheles dirus vitellogenin gene and expression and purification of its functional domain
- 第三军医大学学报 2021年43卷第12期页码：1140-1145
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)军事预防医学系热带医学教研室
  2.510220 广州，中国人民解放军陆军第七十四集团军医院重症医学科
- 作者简介：
  
  [王英，电话: (023)68771576，E-mail: wangyingtmmu@126.com WANG Ying, E-mail: wangyingtmmu@126.com
- 基金信息：
  
  重庆市自然科学基金项目(cstc2018jcyjAX0182);国家自然科学基金(81702035);国家自然科学基金(81971971)
- DOI：10.16016/j.1000-5404.202012233
  中图分类号： Q962;Q963;Q966
- 收稿：2020-12-29，
  
  修回：2021-02-02，
  
  纸质出版：2021-06-30
- 稿件说明：
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于莎莎, 胡辉, 秦杰, 等. 大劣按蚊卵黄蛋白原基因cDNA全长序列扩增及功能域蛋白的表达与纯化研究[J]. 第三军医大学学报, 2021,43(12):1140-1145.

Shasha YU, Hui HU, Jie QIN, et al. Amplification of full-length cDNA sequence of Anopheles dirus vitellogenin gene and expression and purification of its functional domain[J]. Journal of Third Military Medical University, 2021, 43(12): 1140-1145.
于莎莎, 胡辉, 秦杰, 等. 大劣按蚊卵黄蛋白原基因cDNA全长序列扩增及功能域蛋白的表达与纯化研究[J]. 第三军医大学学报, 2021,43(12):1140-1145. DOI： 10.16016/j.1000-5404.202012233.

Shasha YU, Hui HU, Jie QIN, et al. Amplification of full-length cDNA sequence of Anopheles dirus vitellogenin gene and expression and purification of its functional domain[J]. Journal of Third Military Medical University, 2021, 43(12): 1140-1145. DOI： 10.16016/j.1000-5404.202012233.

摘要

目的

对大劣按蚊卵黄蛋白原(

Anopheles dirus

vitellogenin

AdVg)基因cDNA的全长序列进行扩增和分析，并对其功能域蛋白进行表达与纯化。

方法

基于前期克隆的AdVg基因cDNA片段，利用RACE PCR技术，扩增AdVg基因的5'末端及3'末端序列。用Vector NTI软件将测序得到的AdVg cDNA片段、5'末端序列、3'末端序列进行校正，拼接得到完整的AdVg cDNA全长序列。对AdVg全长序列进行功能域预测。利用pet28a(+)构建重组质粒进行AdVg功能域蛋白的表达与纯化。

结果

成功获得AdVg基因全长cDNA序列，包括117 bp的5' UTR序列、192 bp的3' UTR序列和6 186 bp的ORF序列。AdVg蛋白主要包含Vit N、DUF1943及vWD 3个结构域。通过构建重组质粒，成功表达和纯化了AdVg Vit N功能域蛋白。

结论

成功克隆了大劣按蚊卵黄蛋白原基因的全长序列，并获得了重组蛋白。

Abstract

Objective

To amplify and analyze the full-length cDNA of

Anopheles dirus

vitellogenin (AdVg) gene

and express and purify its functional domain.

Methods

Based on the cDNA fragments of AdVg gene cloned earlier

the full-length sequence was obtained through rapid amplification of cDNA ends (RACE). The sequences of AdVg cDNA fragment

5' end and 3' end were corrected and assembled to obtain a complete AdVg cDNA full-length sequence by Vector NTI software. Then the full-length sequence of AdVg was used to predict the functional domain. Finally

recombinant plasmid pet28A (+) was used to express and purify the AdVg functional domain.

Results

The full-length cDNA sequence of the AdVg gene was successfully obtained

including a 117 bp 5' UTR sequence

a 192 bp 3'UTR sequence

and a 6 186 bp open reading frame (ORF) sequence. Meanwhile

the analysis showed that the AdVg protein mainly contained 3 structural domains

including Vit N

DUF1943 and vWD. Then we successfully expressed and purified the functional domain AdVg Vit N by constructing a recombinant plasmid.

Conclusion

The full-length cDNA of AdVg is successfully cloned

and the recombinant protein of its functional domain Vit N is obtained in this study.

关键词

Keywords

references

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