CAJ | 학술논문

运用了抑制性差减杂交和cDNA芯片技术鉴定中华绒鳌蟹卵巢发育相关基因,并用5'和3'RACE的方法克隆了一个与Ⅱ期相比在卵巢发育的Ⅲ期高表达的新基因EJO3(GenBank登录号:AY185919).EJO3基因的开放阅读框(ORF)长1 368 bp,编码455个氨基酸.从氨基酸序列推算的EJO3的等电点和相对分子质量分别为5.79和50 000.生物信息学分析表明,EJO3可能是一个含有VWD结构域的分泌蛋白.EJO3在Ⅱ期和Ⅲ期卵巢的差异表达用Northern blot进行了进一步验证.组织表达谱分析表明EJO3在卵巢高表达,在肠、心脏、肌肉和肝胰腺中弱表达或不表达.本研究结果为进一步深入研究中华绒鳌蟹卵巢发育的分子机制奠定了重要的前期工作基础.
운용료억제성차감잡교화cDNA심편기술감정중화융오해란소발육상관기인,병용5'화3'RACE적방법극륭료일개여Ⅱ기상비재란소발육적Ⅲ기고표체적신기인EJO3(GenBank등록호:AY185919).EJO3기인적개방열독광(ORF)장1 368 bp,편마455개안기산.종안기산서렬추산적EJO3적등전점화상대분자질량분별위5.79화50 000.생물신식학분석표명,EJO3가능시일개함유VWD결구역적분비단백.EJO3재Ⅱ기화Ⅲ기란소적차이표체용Northern blot진행료진일보험증.조직표체보분석표명EJO3재란소고표체,재장、심장、기육화간이선중약표체혹불표체.본연구결과위진일보심입연구중화융오해란소발육적분자궤제전정료중요적전기공작기출.
In the present study, we performed suppression subtractive hybridization (SSH) combining cDNA macroarray to identify genes putatively involved in the ovarian development of Chinese mitten crab. A novel gene, EJO3 (GenBank accession number; AY 185919) whose expression is higher at stage III than that at stage II was cloned by 5' and 3' RACE. The ORF (open reading frame, ORF) of the cDNA of EJ03 consisted of 1 368 bp encoding 455 amino acids. The pI/Mw deduced from the amino acids sequence was 5.79/50 000. Protein coded by EJO3 gene was probably a secretion protein with a VWD domain. The differential expression of EJO3 was further confirmed by Northern blot analysis. Expression profile analysis showed that EJ03 is highly expressed in ovary and there is little or no expression of EJO3 in intestine, heart, muscle and hepatopancreas. Results of this study have made an important foundation to further elucidate the molecular mechanism of the ovarian development of Chinese mitten crab.