小菜蛾<i>COX</i>7<i>C</i>基因的克隆与表达分析

陈伟军; 谢苗; 仲帆; 尤燕春; 尤民生

doi:10.19303/j.issn.1008-0384.2017.04.009

小菜蛾COX7C基因的克隆与表达分析

doi: 10.19303/j.issn.1008-0384.2017.04.009

陈伟军^{1, 2, 3, 4,},
谢苗^{1, 2, 3, 4},
仲帆²,
尤燕春^{1, 2, 3, 4},
尤民生^{1, 3, 4, ,}

1.
福建农林大学应用生态研究所, 福建福州 350002
2.
福建农林大学生命科学学院, 福建福州 350002
3.
农业部闽台作物有害生物综合治理重点实验室, 福建福州 350002
4.
闽台特色作物病虫生态防控协同创新中心, 福建福州 350002

基金项目:

国家自然科学基金项目 31230061

国家自然科学基金项目 31320103922

详细信息

作者简介:
陈伟军(1991-), 男, 硕士研究生, 研究方向:生物化学与分子生物学(E-mail:411585662@qq.com)

通讯作者:
尤民生(1954-), 男, 博士, 教授, 研究方向:害虫生态控制(E-mail:msyou@iae.fjau.edu.cn)

中图分类号: S435.132
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- 被引次数: 0
出版历程
- 收稿日期: 2017-02-06
- 修回日期: 2017-03-15
- 刊出日期: 2017-04-28

Characterization and Expression Profiling of COX7C genes in Plutella xylostella L

CHEN Wei-jun^{1, 2, 3, 4
,},
XIE Miao^{1, 2, 3, 4},
ZHONG Fan²,
YOU Yan-chun^{1, 2, 3, 4},
YOU Min-sheng^{1, 3, 4
, ,}

1.
Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China
2.
College of Life Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China
3.
Fujian-Taiwan Joint Centre for Ecological Control Crop Pests, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China
4.
Key Laboratory of Integrated Pests Management for Fujian-Taiwan Crops, Ministry of Agriculture, Fuzhou, Fujian 350002, China

摘要

摘要: 细胞色素C氧化酶（Cytochrome C oxidase，COX）是真核细胞线粒体内膜和需氧细菌细胞膜呼吸电子传递链的终端氧化酶，与ATP的合成密切相关，COX7C基因属于细胞色素C氧化酶核基因编码的一个亚基。本研究克隆了小菜蛾COX7C基因，测序后获得1条全长为213 bp的PxCOX7C基因开放阅读框序列，进化树分析表明COX7C在同一目昆虫中也不是完全保守的。荧光定量PCR分析COX7C在小菜蛾毒死蜱品系（CRS）、氟虫腈品系（FRS）和敏感品系（SS）3龄幼虫中的表达水平，结果表明COX7C在3个品系中均有表达，且在FRS中的表达量显著高于SS，说明其可能与小菜蛾对氟虫腈解毒代谢相关。本研究对小菜蛾COX7C进行了初步的探索研究，为今后对COX7C进一步的功能研究奠定基础。
- 小菜蛾 /
- 细胞色素C氧化酶 /
- 抗药性 /
- qPCR
Abstract: Cytochrome C oxidase (COX) is a member of the heme-copper oxidase superfamily, which is a terminal oxidase of eukaryotic mitochondrial intima and aerobic bacterial cell membrane respiratory electron transport chain, and is closely related to the synthesis of ATP. The COX7C gene is a subunit of the cytochrome C oxidase. In this study, the open reading frame COX7C gene of Plutella xylostella was cloned and sequenced, which includes 213 bp nucleotide. The phylogenetic tree analysis showed that COX7C was not highly conserved between the same order insects. The expression levels of COX7C in the third instar larvae of chlorpyrifos-, fipronil-resistant and susceptible strains were analyzed by qPCR. The results showed that COX7C was expressed in all three strains, and the expression level in the fipronil-resistant strain was significantly higher than that in SS, indicating that it might be involved in the process of detoxification of fipronil in P. xylostella. This study only conducted a preliminary study of COX7C in the P.xylostella, and a foundation for further research on the function of COX7C in the future.
- Plutella xylostella /
- Cytochrome Coxidase /
- Resistance /
- qPCR

HTML全文

图 1 PxCOX7C的核苷酸序列及其推导的氨基酸序列

Figure 1. Nucleotide sequence and deduced amino acid sequence of PxCOX7C

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图 2 COX7C在不同昆虫中的系统进化树

Figure 2. Phylogenetic tree of COX7C in different species of insects

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图 3 PxCOX7C在不同品系中的RPKM值

Figure 3. RPKM values of PxCOX7C in different strains

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图 4 PxCOX7C在不同品系中的相对表达量

注：图中所有数据均为3次平行试验的相对表达量(均值±SE)，表达量计算采用2^-△Ct法。不同字母表示差异显著(Tukey's test, P＜0.05)。

Figure 4. Expression level of the PxCOX7C at different strains based on qPCR

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表 1 目的基因引物

Table 1. Primer sequences used in the experiments

名称	引物序列(5'-3')	产物长度/bp	引物信息
COX7C	GGAATTCCATATGATGCACGCCCCACTCGCT CCGCTCGAGTTACTTCTTAAGCAGCTGGTGGCG	213	cDNA扩增上游引物 cDNA扩增下游引物
qPCR-COX7C	ACCTGCCGTTCGACATCAAC TGGTGGCGAGTGATGAGGTA	150	qPCR上游引物 qPCR下游引物
RPS4	ATGGATGTTGTGTCGATTGAAAAGA GAGTGATGCGGTGGATGGTGA	88	内参基因上游引物内参基因下游引物

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