摘要
为了探究亲环蛋白B(Cy PB)在马氏珠母贝中的作用机制,运用c DNA末端快速扩增(RACE)技术克隆得到马氏珠母贝Cy PB基因(Pm Cy PB)c DNA的全长序列,并且应用实时荧光定量PCR技术对Pm Cy PB基因在马氏珠母贝不同组织中的表达进行检测。结果显示,Pm Cy PB基因序列全长1266 bp,其中开放阅读框(ORF)为678 bp,编码225个氨基酸,5′-UTR为62 bp,3′-UTR为526 bp。预测其相对分子量为24829.3,理论等电点5.77。多序列比对结果显示Pm Cy PB基因与其他物种的Cy PB具有较高的保守性。SMART软件对Pm Cy PB进行蛋白质序列分析,发现它包含亲环蛋白家族典型的肽脯氨酰顺反异构酶的结构域。实时荧光定量PCR数据分析表明,该Pm Cy PB基因在马氏珠母贝闭壳肌、肝胰腺、血细胞、外套膜、足、性腺、鳃这7种组织中均有表达,在外套膜表达量最高,其次是鳃。结果可为进一步阐述Pm Cy PB在马氏珠母贝中的免疫防御机制提供一定的理论基础。
In order to explore the action mechanism of cyclophilin B in Pinctada martensii, the full length of Pm Cy PB gene were obtained using rapid amplification technology of c DNA ends. And expression of Pm Cy PB in different tissues was tested by Real-time PCR. The results showed that the total length of Cy PB gene was1266 bp, including 678 bp of the open reading frame（ORF） which encoding 225 amino acids, a 5-＇UTR of 62 bp and a 3＇-UTR of 526 bp. The predicted molecular weight was 24829.3, and the isoelectric point was 5.77.Multiple sequence alignment showed that Pm Cy PB was highly conservative in Cy PB gene with other species.By SMART software analysis, we found that it contained the typical domain of the Cy PB family, namelypeptidylcis-transisomerase. In addition, real-time PCR indicated that Pm Cy PB could be expressed in all ofthe detected tissues, including adductor muscle, hepatopancreas, haemocytes, mantle, foot, gill, gonads, withthe highest expression in mantle and the second in gill. The results could provide certain theoretical basis forthe further study of Pm Cy PB in the immune response in mollusk.
出处
《中国农学通报》
2015年第8期57-63,共7页
Chinese Agricultural Science Bulletin
基金
国家自然科学基金"马氏珠母贝珍珠质形成相关micro RNA的鉴定与功能研究"(41206141)
国家自然科学基金"基于珍珠贝基因组测序分析的珍珠形成矿化关键基因和蛋白质的研究"(31272635)
国家自然科学基金"马氏珠母贝生长性状QTLs精细定位与相关基因功能分析"(31372526)
关键词
马氏珠母贝
CY
PB
基因克隆
荧光定量
表达分析
Pinctada martensii
Cy PB
gene cloning
fluorescent quantitation
expression analysis
