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Molecular cloning and characterization of SoxB2 gene from Zhikong scallop Chlamys farreri 被引量:1

Molecular cloning and characterization of SoxB2 gene from Zhikong scallop Chlamys farreri
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摘要 The Sox proteins play critical roles during the development of animals, including sex determination and central nervous system development. In this study, the SoxB2 gene was cloned from a mollusk, the Zhikong scallop (Chlamysfarreri), and characterized with respect to phylogeny and tissue distribution. The full-length cDNA and genomic DNA sequences of C. farreri SoxB2 (CflSoxB2) were obtained by rapid amplification of cDNA ends and genome walking, respectively, using a partial cDNA fragment from the highly conserved DNA-binding domain, i.e., the High Mobility Group (HMG) box. The full-length cDNA sequence of CJSoxB2 was 2 048 bp and encoded 268 amino acids protein. The genomic sequence was 5 551 bp in length with only one exon. Several conserved elements, such as the TATA-box, GC-box, CAAT-box, GATA-box, and Sox/sry-sex/testis-determining and related HMG box factors, were found in the promoter region. Furthermore, real-time quantitative reverse transcription PCR assays were carried out to assess the mRNA expression of CJSoxB2 in different tissues. SoxB2 was highly expressed in the mantle, moderately in the digestive gland and gill, and weakly expressed in the gonad, kidney and adductor muscle. In male and female gonads at different developmental stages of reproduction, the expression levels of CfS;oxB2 were similar. Considering the specific expression and roles of SoxB2 in other animals, in particular vertebrates, and the fact that there are many pallial nerves in the mantle, cerebral ganglia in the digestive gland and gill nerves in gill, we propose a possible essential role in nervous tissue function for SoxB2 in C.farreri. The Sox proteins play critical roles during the development of animals,including sex determination and central nervous system development.In this study,the SoxB2 gene was cloned from a mollusk,the Zhikong scallop(Chlamys farreri),and characterized with respect to phylogeny and tissue distribution.The full-length cDNA and genomic DNA sequences of C.farreri SoxB2(CfSoxB2) were obtained by rapid amplification of cDNA ends and genome walking,respectively,using a partial cDNA fragment from the highly conserved DNA-binding domain,i.e.,the High Mobility Group(HMG) box.The full-length cDNA sequence of CfSoxB2 was 2 048 bp and encoded 268 amino acids protein.The genomic sequence was 5 551 bp in length with only one exon.Several conserved elements,such as the TATA-box,GC-box,CAAT-box,GATA-box,and Sox/sry-sex/testis-determining and related HMG box factors,were found in the promoter region.Furthermore,real-time quantitative reverse transcription PCR assays were carried out to assess the mRNA expression of CfSoxB2 in different tissues.SoxB2 was highly expressed in the mantle,moderately in the digestive gland and gill,and weakly expressed in the gonad,kidney and adductor muscle.In male and female gonads at different developmental stages of reproduction,the expression levels of CfSoxB2 were similar.Considering the specific expression and roles of SoxB2 in other animals,in particular vertebrates,and the fact that there are many pallial nerves in the mantle,cerebral ganglia in the digestive gland and gill nerves in gill,we propose a possible essential role in nervous tissue function for SoxB2 in C.farreri.
出处 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2013年第6期1216-1225,共10页 中国海洋湖沼学报(英文版)
基金 Supported by the National Natural Science Foundation of China(Nos.31072190,30972239) the National Basic Research Program of China(973 Program)(No.2010CB126406) the Taishan Scholar Program of Shandong Province the Doctoral Fund of Ministry of Education of China(No.20100132110014) the Natural Science Foundation of Shandong Province(No.ZR2009DM019)
关键词 SoxB2 gene organization PROMOTER QRT-PCR Chlamysfarreri 基因组序列 栉孔扇贝 分子克隆 全长cDNA序列 中枢神经系统 逆转录聚合酶链反应 DNA结合结构域 鉴定
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