摘要
【目的】克隆野桑蚕（Bombyx mandaina）核糖体蛋白L21（RPL21）基因并进行半定量分析,为研究RPL21基因在昆虫体内的生物学功能提供参考。【方法】采用RT-PCR克隆野桑蚕RPL21基因序列,并采用半定量PCR分析该基因在野桑蚕4龄幼虫不同组织中的表达情况。【结果】扩增获得的野桑蚕RPL21基因序列全长为586 bp,其包含一个完整的开放阅读框（ORF）,长度为480 bp,编码159个氨基酸;预测蛋白的分子量和等电点分别为18 k Da和11.01。序列比对结果表明,野桑蚕与其他12个物种的RPL21基因的相似性介于55.5%-91.6%,其中与致倦库蚊（Culex quinquefasciatus）RPL21的相似性最低,与家蚕（Bombyx mori）的相似性最高。半定量分析结果表明,野桑蚕RPL21基因在野桑蚕幼虫的中肠、丝腺、脂肪体、血液、睾丸、表皮、卵巢和脑组织中均有表达,且转录水平无明显差异。【结论】野桑蚕RPL21基因进化较保守,在各组织中分布广泛且表达水平稳定。
【Objective 】The cloning and sequencing analysis of ribosomal protein L21（RPL21） gene from Bombyx mandaina was conducted in the present study, as well its expression in different tissues, in order to provide references for researching its biological function in insects. 【Method】The RPL21 gene was cloned from fat body tissues of larvae by RTPCR. The transcriptional levels of RPL21 gene in midgut, silk gland, fat body, hemocytes, testis, integument, ovary and brain of fourth instar larvae on the 3rdday were analyzed by using semi-quantitative PCR. 【Result】The full-length c DNA of RPL21 was 586 bp in length, which contained an open reading frame（ORF） of 480 bp encoding a deduced polypeptide of 271 amino acids; and the theoretical molecular weight of 18 k Da and isolectric point of 11.01 were predicted for RPL21 protein. The results of sequence alignment revealed that, the similarity of B. mandaina RPL21 gene with other 12 species ranged from 55.5% to 91.6%, the highest was found with Bombyx mori and the lowest with Culex quinquefasciatus, respectively. The results of semi-quantitative PCR analysis showed that, the B. mandaina RPL21 gene could be extensively expressed in mid-intestine, silk gland, fat body, hemocytes, testis, integument, ovary and brain of larvae, and no significant difference was found in their transcriptional levels. 【Conclusion】The evolution of B. mandaina RPL21 gene was conservative; RPL21 gene is widely distributed in different tissues with a stable transcriptional level.
出处
《南方农业学报》
CAS
CSCD
北大核心
2016年第2期290-295,共6页
Journal of Southern Agriculture
基金
supported by Natural Science Foundation of Jiangsu Province(13KJB180010)
