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利用荧光差异显示技术分离的家蚕抗NPV相关基因s3a 被引量:13

Fluorescent differential display analysis of the gene s3a related to NPV resistance in Bombyx mori L.
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摘要 通过荧光差异显示技术,分析了家蚕Bombyxmori对BmNPV抗性品系NB、感性品系306和近等基因系306NNZZ添毒和未添毒处理区的基因表达的差异。根据差异显示的结果克隆了一条702bp长度的cDNA片段,并用Northernblot进行了验证。该序列经过NCBIEST库的同源性比较获得了电子延伸。延伸后的序列用特异引物进行RT_PCR扩增获得了一条782bp的序列,拼接后基因cDNA序列全长为827bp,推导的氨基酸序列与草地夜蛾SpodopterafrugiperdaS3A同源性最高达97.7%;其次是烟芽夜蛾HeliothisvirescensS3A,同源性为94.0%;与黑腹果蝇DrosophilamelanogasterS3A同源性为75.3%。比较结果显示这是一个新的家蚕基因,定名为家蚕s3a基因。本实验获得的s3a基因在家蚕感性和抗性品系以及添毒处理和未添毒处理中都具有差异表达,其中在抗性品系和近等基因系中的表达高于感性品系,在添毒处理中的表达高于未添毒组。因此推测它是一个与家蚕抗BmNPV相关的新基因。 Using fluorescent differential display (FDD) technique, the differential expression of genes related to BmNPV resistance in highly resistant strain NB, highly susceptible strain 306 and near isogenic line 306NNZZ of the silkworm, Bombyx mori L. was analyzed. Based on the differential display bands, a 702 bp fragment named C18 (702) was cloned and confirmed by Northern blot hybridizations. The sequence was then electrically extended based on homology comparison with NCBI ESTs, and further confirmed by RT_PCR using specific primers. A novel gene was characterized and revealed to encode a putative BmS3A protein. The gene had 97.7% homology to Spodoptera frugiperda S3A, 94% homology to Heliothis virescens S3A and 75.3% homology to Drosophila melanogaster S3A, and was named B. mori s3a (Bms3a). It had higher expression in the highly resistant strain and near isogenic line than in the highly susceptible silkworm strain, and it also had higher expression in BmNPV treated strains than untreated strains. The results suggested that Bms3a is involved in silkworm BmNPV resistance.
出处 《昆虫学报》 CAS CSCD 北大核心 2005年第3期347-352,共6页 Acta Entomologica Sinica
基金 国家自然科学基金项目(30370773) 江苏大学高级人才基金(1291180009)
关键词 家蚕 BMNPV 基因 s3a 抗性 荧光差异显示 Bombyx mori BmNPV gene s3a resistance fluorescent differential display
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