褐飞虱Nilaparvata lugens(S"非汉字符号"l)肌动蛋白基因3′-RACE及基因表达的RT-PCR检测

3′-RACE of actin genes in Nilaparvata lugens (S■l)and RT-PCR examination on expression of the actin genes

通过锚定的 3′ RACE筛选实验 ,确定锚定效率最好的下游引物 ,用于褐飞虱各发育期肌动蛋白基因表达的RT PCR检测。结果表明 :3个锚定引物中 ,0 4 2 2 7( 5′ >TCACACAGGAAACAGCTATGACTTTTTTTTTTTTTTA <3′)的锚定效率最好 ,可以扩增出 5条褐飞虱的肌动蛋白基因 3′末端片段 ,依其大小命名为BPH A、BPH B、BPH C、BPH D、BPH E。RT PCR检测表明 :BPH A从 3龄开始到成虫期都有常量表达 ;BPH B、BPH C、BPH E从 2龄开始到成虫期都有表达 ;BPH D在整个幼虫期都有表达 。

The most efficient anchor primer decided by tentative 3′ RACE,0422 7(5′>TCA CAC AGG AAA CAG CTA TGA CTTTTTTTTTTTTTT A<3′),was applied to exam expression of the actin genes in the brown planthopper, Nilaparvata lugens (Stl).At least five DNA fragments were amplified from the actin gene 3′ terminals,meaning that there are more than five actin genes in Nilaparvata lugens (Stl).Examination for the expression of actin genes showed that BPH A can be found from the 3rd instar nymph,BPH D in nymphal stage only,and BPH B,C,E in all stages from the 2nd instar nymph.