血清反应因子(SRF)是一个高度保守的转录因子,在细胞增殖、细胞凋亡以及免疫反应中发挥重要作用。为了探究血清反应因子在马氏珠母贝中的生物学功能,本研究运用cDNA末端快速扩增(RACE)技术克隆得到马氏珠母贝SRF基因(PmSRF)cDNA的全长序列,并且应用实时荧光定量PCR技术对PmSRF基因在马氏珠母贝不同组织中的表达进行检测。结果显示,PmSRF基因序列全长1758 bp,其中开放阅读框(ORF)为1440 bp,编码479个氨基酸,5'UTR为65 bp,3'UTR为253 bp,包含29 bp的polyA。预测其相对分子量为50 534.6 Da,理论等电点为7.71。多序列比对结果发现物种间SRF具有较高的保守性。SMART软件分析显示PmSRF具有典型的MADS结构域。荧光定量PCR数据分析表明,PmSRF基因在马氏珠母贝闭壳肌、肝胰腺、血细胞、外套膜、性腺、鳃六种组织中均有表达,其中在鳃中表达量最高。本研究可为进一步探究PmSRF在贝类中的生物学功能提供重要的理论基础和参考价值。
Serum response factor(SRF) is a highly conservative transcription factor, and plays an important role in cell proliferation, apoptosis, and immune response. In order to explore the biological function of SRF in Pinctada martensii(PmSRF), the full length of PmSRF gene was obtained using rapid amplification of cDNA ends technology. And expression of PmSRF in different tissues was tested by real-time PCR. The results showed that the total length of PmSRF gene was 1 758 bp, including 1 440 bp of the open reading frame(ORF) which encodes 479 amino acids, a 5' UTR of 65 bp and a 3' UTR of 253 bp with 29 bp polyA. The predicted molecular weight is50 534.6 Da, and the isoelectric point is 7.71. Multiple sequence alignment showed that SRF was highly conservative among species. Protein sequence analysis showed that PmSRF has a typical MADS structural domain. In addition,real-time PCR analysis showed PmSRF could be detected in all of tested tissues, including adductor muscle,hepatopancreas, haemocytes, mantle, gill and gonads, with the highest expression level in the gill. Our results could provide basis for the further study of the biological function of PmSRF in the shellfish.