小菜蛾γ- 氨基丁酸受体基因片段的克隆、表达及特征分析

园艺学报 ›› 2006, Vol. 33 ›› Issue (2): 300-305.

小菜蛾γ- 氨基丁酸受体基因片段的克隆、表达及特征分析

周小毛^{1, 2, 3};吴青君^1*;胡美英²;张友军¹;朱国仁¹;徐宝云¹

(¹中国农业科学院蔬菜花卉研究所, 北京100081; ² 华南农业大学昆虫毒理研究室, 广州510642; ³湖南农业大学农药学系, 长沙410128)

收稿日期:2005-07-20 修回日期:2005-10-09 出版日期:2006-04-25 发布日期:2006-04-25

Cloning and Expression Characteristics of cDNA Fragment of GABAa Receptor Gene in Diamondback Moth ( Plutella xylostella )

Zhou Xiaomao^{1, 2, 3};Wu Qingjun^1*;Hu Meiying²;Zhang Youjun¹;Zhu Guoren¹; Xu Baoyun¹

(¹Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China; ²Laboratory of Insect Toxicology, South China Agricultural University, Guangzhou 510642, China; ³Department of Pesticide Science, Hunan Agricultural University, Changsha 410128, China)

Received:2005-07-20 Revised:2005-10-09 Online:2006-04-25 Published:2006-04-25

摘要/Abstract

摘要： 利用反转录多聚酶链式反应(RT-PCR) 对小菜蛾( Plutella xylostella ) 的γ - 氨基丁酸a(GABAa) 受体基因cDNA片段进行了克隆和序列分析。通过简并性上游引物和下游引物扩增出小菜蛾GABAa受体基因248 bp的cDNA片段。该cDNA基因片段的氨基酸与已报道的烟芽夜蛾(Heliothis viresce)的GABAa受体基因氨基酸序列同源性为100%。以GABAa受体基因片段为基础, 构建了优化的半定量RT-PCR法, 以18S rRNA为内标, 研究小菜蛾不同组织以及不同发育阶段GABAa受体基因表达的差异。结果表明, GABAa受体基因在头部表达量最高, 胸部次之, 腹部最低; 在不同发育阶段的表达量差异不大。

关键词: 小菜蛾, γ - 氨基丁酸受体基因, 半定量RT-PCR, 基因表达

Abstract: One fragment of 248 bp cDNA of the GABAa recep tor gene from the fourth instar larvae of diamondback moth (DBM, Plutella xylostella) was amplified, cloned, sequenced and analyzed by using reverse transcription polymerase chain reaction (RT-PCR) method. Homologous analysis showed that it had 100% homologywith the published deduced amino acid sequences of Helioth is viresce. Based on the GABAa receptor gene clone, an optimal emiquantitative RT-PCR method was successfully constructed. Using 18S rRNA as internal control, the exp ression difference of GABAa recep tor gene in different tissues and different developmental stages was studied. Results showed that the gene was expressed the highest level in heads of DBM, next was in thorax and the lowest level in abdomen. The exp ression at different developmental stages was not significantly different.

Key words: Plutella xylostella, GABAa receptor gene, Semi-quantitative RTPCR, Gene expression

周小毛;;;吴青君;胡美英;张友军;朱国仁;徐宝云. 小菜蛾γ- 氨基丁酸受体基因片段的克隆、表达及特征分析[J]. 园艺学报, 2006, 33(2): 300-305.

Zhou Xiaomao;;;Wu Qingjun;Hu Meiying;Zhang Youjun;Zhu Guoren;Xu Baoyun. Cloning and Expression Characteristics of cDNA Fragment of GABAa Receptor Gene in Diamondback Moth ( Plutella xylostella )[J]. ACTA HORTICULTURAE SINICA, 2006, 33(2): 300-305.

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