棉铃虫细胞色素P450CYP6B6基因启动子的活性分析

李芬; 马纪; 刘小宁

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棉铃虫细胞色素P450CYP6B6基因启动子的活性分析

更新时间：2026-01-21

- 棉铃虫细胞色素P450CYP6B6基因启动子的活性分析
- Journal of Xinjiang University (Natural Science Edition in Chinese and English) Vol. 29, Issue 1, Pages: 13-18(2012)
- 作者机构：
  
  新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室
- 作者简介：
- 基金信息：
- DOI：
  CLC： S435.622.3
- Published：2012
- 稿件说明：
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[1]李芬,马纪,刘小宁.棉铃虫细胞色素P450CYP6B6基因启动子的活性分析[J].新疆大学学报(自然科学版),2012,29(01):13-18.

李芬, 马纪, 刘小宁. 棉铃虫细胞色素P450CYP6B6基因启动子的活性分析[J]. Journal of Xinjiang University (Natural Science Edition in Chinese and English), 2012, 29(1): 13-18.
[1]李芬,马纪,刘小宁.棉铃虫细胞色素P450CYP6B6基因启动子的活性分析[J].新疆大学学报(自然科学版),2012,29(01):13-18. DOI：

李芬, 马纪, 刘小宁. 棉铃虫细胞色素P450CYP6B6基因启动子的活性分析[J]. Journal of Xinjiang University (Natural Science Edition in Chinese and English), 2012, 29(1): 13-18. DOI：

摘要

在昆虫细胞色素P450s超家族中

CYP6家族基因与杀虫剂抗性关系最为密切

其中CYP6B家族基因的表达在植物次生物质的代谢调节中发挥重要作用.为研究CYP6B6基因的表达调节机制

本文采用PCR的方法从棉铃虫中肠DNA序列中扩增了CYP6B6基因转录起始位点上游不同片段长度的5段序列

构建由其驱动的萤火虫荧光素酶基因报告质粒pGL3-CYP6B6-promoter

与内参载体pRL-TK同在脂质体的介导下转染昆虫Sf9细胞

体外分析该基因启动子的转录活性及与调控相关的因素.结果显示

5个报告质粒的荧光素酶相对活性分别是基本载体pGL3-Basic载体的0.95、1.95、1.21、1.17和1.01倍

其中p-791片段序列具有启动子活性

这段序列位于翻译起始位点上游400bp之间

生物信息学分析显示

该区间具有TATA box、CAAT box、CBFα、C/EBP等基本元件和AhR等异源物质响应的重要元件结合序列

暗示该序列可能具有与转录调控相关的元件

这为研究CYP6B6的调控机制奠定了基础.

Abstract

In the cytochrome P450s superfamily of insects

CYP6 family genes are closely associated with insecticide resistance

which CYP6B family gene’s expression plays an important role involved in the catabolism and anabolism of plant secondary compounds.To evaluate the regulatory mechanism of CYP6B6 transcription

five different length fragments from the upstream of CYP6B6 transcriptional start site was isolated by PCR from Helicoverpa amigera midgut

and then the five promoters are cloned to pGL3 vector with luciferase report gene.Through the liposome mediating co-transfect Sf9 cells together with pRLTK vector

we analyse transcription activity and regulation related factors of the promoters in vitro.The results showed that the relative luciferase activity of five promoters were 0.95

1.95

1.21

1.17 and 1.01fold

respectively

compared with pGL3-Basic plasmid.The fragment p-791 showed promoter activity.The bioinformatics analysis indicated that this fragment about 400bp upstream of ATG has binding sites for basic transcriptional factors including TATA box

CAAT box

CBFα

C/EBP and xenobiotics responsing element AhR

implying that this sequence may have some elements relative to transcription regulation

which lays the foundation for the studies of CYP6B6 regulatory mechanism.

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Keywords

references

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