摘要
采用c DNA末端快速扩增技术克隆获得马氏珠母贝清道夫受体基因c DNA全长序列(Pm SR-B);利用荧光定量技术检测Pm SR-B基因在各个组织中的表达量。结果表明,Pm SR-B基因c DNA序列全长1 857 bp,开放阅读框(ORF)长1 443 bp,编码480个氨基酸,5′非翻译区(5′UTR)长89 bp,3′UTR长325 bp。预测其分子质量为54.77 ku,等电点为7.94,脂溶性系数92.94,总平均亲水性-0.120,属于疏水性蛋白;不稳定指数26.37,属于稳定蛋白。氨基酸序列同源比对结果,Pm SR-B氨基酸序列与其他物种具有一定的保守性,与华贵类栉孔扇贝(Mimachlamys nobilis)SR-B的序列的相似度高达47%。荧光定量PCR检测结果,Pm SR-B在肝胰腺中表达量最高,其后依次是鳃、闭壳肌、中央膜,各组织的表达量差异具统计学意义(P<0.05)。
The full length of pearl oyster Pinctada fucata martensii scavenger receptor B (PmSR-B) was obtained using rapid amplification of cDNA ends technology. The expression of tissue-specific was detected by real-time PCR. The results showed that the total length of PmSR-B gene was 1 857 bp,including 1 443 bp of the open reading frame (ORF) that encoded 480 amino acids, a 5′ UTR of 89 bp and a 3′ UTR of 325 bp. The predicted molecular weight was54.77 ku and isoelectric point was 7.94.The Aliphatic index was 92.94 and the Grand average of hydropathicity (GRAVY) was -0.493. The instability index was 26.37, which belongs to the stable protein. Multiple sequence alignment indicated that PmSR-B was some extent conservative among species,with high identity with Mimachlamys nobilis. Fluorescence quantitative PCR analysis showed that the hepatopancreas had the highest expression of PmSR-B gene, followed by gill, adductor muscle and central membrane.
作者
雷超
贝伟烈
郑哲
罗少杰
王庆恒
邓岳文
LEI Chao;BEI Wei-lie;ZHENG Zhe;LUO Shao-jie;WANG Qing-heng;DENG Yue-wen(Fisheries College, Guangdong Ocean University;Pearl Breeding and Processing Engineering Technology Research Center of Guangdong Province, Zhanjiang 524088, China)
出处
《广东海洋大学学报》
CAS
2017年第1期7-14,共8页
Journal of Guangdong Ocean University
基金
广东省海洋与渔业局科技专项(A201308A10)
2015年广东海洋大学"海之帆-起航计划"大学生科技创新项目
2015年广东海洋大学水产学院创新项目
关键词
马氏珠母贝
清道夫受体B
基因克隆
基因表达
Pinctada fucata martensii
scavenger receptor B
Gene cloning
Gene expression
