褐飞虱NADH泛醌氧化还原酶51kDa亚基cDNA片段的克隆及表达分析

Cloning and Expression of NADH-quinone Oxidoreductase 51kDa Subunit Gene from Brown Planthopper Nilaparvata Lugens

NADH泛醌氧化还原酶是动物体内呼吸链电子传递系统的第一个酶,克隆水稻害虫褐飞虱的NADH泛醌氧化还原酶基因,及研究其在褐飞虱与水稻互作中的表达变化,将为科学防治褐飞虱提供新的线索。利用反转录多聚酶链式反应(RT-PCR)技术克隆了褐飞虱NADH泛醌氧化还原酶51 kDa亚基基因的cDNA片段,并进行了序列测定;使用Northern杂交技术检测了该基因对两种不同抗性水稻的分子反应。分子杂交结果表明,在取食抗性水稻品种B5后,褐飞虱的NADH泛醌氧化还原酶51 kDa亚基基因表达水平明显升高,而取食感虫水稻TN1后,该基因的表达水平没有明显变化。

NADH-quinone oxidoreductase is the first functional enzyme in the respiratory chain in animals. Cloning and expression analysis of this gene in brown planthopper Nilaparvata lugens will benefit the design of scientific programs for controlling the rice pest. A cDNA fragment of NADH-quinone oxidoreductase 51kDa subunit gene from brown planthopper was cloned by RT-PCR in this research. The expression alteration of the gene in response to two different rice varieties was examined by Northern blot hybridization. The results reveal that the length of the cDNA fragment is 538 bp in size, and the amino acid sequence deduced from the cDNA fragment has a high homology with parts of NADH-quinone oxidoreductase 51kDa subunits from Bos taurus, Homo sapiens, Mus musculus, Macaca fascicularis, Leishmania major, Trypanosoma brucei and Neurospora crassa. Northern hybridization analysis shows that the expression of the gene rapidly increases after brown planthopper is fed with resistant rice B5, but there is no expression change of this gene in response to susceptible rice TN1.