与水稻褐飞虱中肠刷状缘膜囊泡(BBMV)相结合的活性短肽筛选

Screening of Peptides bound to Brash Border Membrane Vesicle of Nilaparvata lugens (Hemiptera: Delphacidae)

参照鳞翅目和双翅目昆虫中肠刷状缘膜囊泡(BBMV)的提取方法,成功提取了水稻褐飞虱中肠BBMV蛋白,利用噬菌体短肽文库展示技术,筛选到2个能够与水稻褐飞虱中肠BBMV结合的短肽P1Z和P2Z.以EGFP蛋白为标签通过Pull down和Western blot方法对短肽P1Z、P2Z与褐飞虱中肠BBMV的结合活性进行了验证.结果表明,利用噬菌体短肽文库展示技术,以褐飞虱中肠BBMV作为底物,经过3轮淘洗筛选到的短肽蛋白与褐飞虱中肠BBMV上的潜在受体具有较强的结合能力.通过对短肽结构域的分析发现,短肽结构α两亲区与β两亲区的分布情况是影响短肽与褐飞虱中肠BBMV结合的一个潜在因素.本研究成功筛选并验证了能够与褐飞虱中肠BBMV结合的短肽.

This research aimed to screen peptides which may bind to the brash border membrane vesicle （BBMV） extracted from the gut of rice brown planthopper （Nilaparvata lugens, Hemiptera： Delphacidae）. BBMV of N. lugen was extracted with the same method for extracting BBMV from the gut of lepidopteran and dipteran insects. Then two peptides P1Z and P2Z that may bind to N. lugens gut BBMV were screened by the use of phage display library. Structure and function of screened peptides were analyzed by bio-analysis. Nucleotide of P1Z, P2Z and an unbound peptide UBP were fused to the 5＇ terminus of egfp gene and P1Z/P2Z/UBP-EGFP fusion proteins were prepared. Binding activity of P1Z and P2Z to N. lugens gut BBMV were confirmed by Pull down and Western blot methods. The results showed that by the use of phage display library it is possible to find peptides which can bind to gut BBMV ofN. lugens. Structure and function analysis of screened peptides show that the distribution differences of Alpha amphipathic regions and Beta amphipathic regions are the possible factors to affect BBMV binding activity of peptides. The results of this research could help to develop a new approach to block transmission of rice viruses by N. lugen. Fig 5, Tab 1, Ref 35