摘要
细胞周期蛋白H(cyclin H,cycH)是细胞周期蛋白依赖性激酶活化激酶(CAK)的重要组成元件,在细胞分裂过程中扮演着重要的作用.本研究从拟穴青蟹(Scylla paramamosain)性腺EST文库中筛选出cyc H基因片段,继而克隆出其全长cDNA(Sp-cyc H)序列,并利用qRT-PCR技术检测其在不同组织和性腺不同发育阶段中的差异表达.结果显示,Sp-cycH全长c DNA序列为1 523bp,其中5'UTR为152bp,3'UTR为372bp,ORF为999bp,可编码332个氨基酸,该序列包含典型的cyclin box序列,进一步比对结果显示其属于cyclinH家族.不同组织的qRT-PCR结果显示,Sp-cyc H在卵巢中的表达量最高,与其他组织具有极显著差异(P<0.01);性腺不同发育阶段的表达结果显示,Sp-cycH在O5期卵巢中的表达量显著高于精巢和卵巢的其他各发育时期(P<0.05);在O4和T2、T3时期的表达量亦显著高于O2期和T1期(P<0.05),据此推测Sp-cyc H对拟穴青蟹的性腺发育可能发挥一定作用.
Cyclin H is one of the important elements of cyclin-dependent kinase activating kinase（CAK）, which plays a crucial role in cell division. In the present study, the fragment of cyclin H was selected from the EST library of Scylla paramamosain（named Sp-cyc H）, which was selected to clone its full-length of c DNA sequence by employing SMART RACE technology. The expression level of Sp-cyc H in different tissues in female crab and different gonad development stages were assessed by using realtime PCR technology. The results showed that the full length c DNA of Sp-cyc H was of 1 523 bp, including a 5＇terminal untranslated region（5＇UTR） of 152 bp, a 3＇UTR of 372 bp, and an open reading frame of 999 bp encoding a protein of 332 aa. The protein contained a typical cyclin box series, which belonged to the family of cyclin H. Its expression level in the ovary was extremely higher than other tissues（P 0.01）. Meanwhile, the gonad expression of Sp-cyc H gene in O5 stage was highest among the other stages with significant difference（P〈0.05）; and its level in the O4, T2 and T3 stages were significantly higher than in O2 and T1 stages. These results provided basic data for elucidating the role of Sp-cyc H in the gonad development of the mud crab.
出处
《宁德师范学院学报（自然科学版）》
2016年第2期155-160,共6页
Journal of Ningde Normal University(Natural Science)
基金
国家自然科学基金(31072200
31472266)
集美大学创新团队基金(2010A001)
