CKS1B参与拟穴青蟹性腺发育的研究

doi:10.13560/j.cnki.biotech.bull.1985.2016.11.030

摘要/Abstract

摘要： CDC28蛋白激酶调节亚基1B（CDC28 protein kinase regulatory subunit 1B,CKS1B）是高度保守的CKS1家族成员之一,在细胞周期中作为细胞周期蛋白激酶的调节亚基参与调控真核生物的细胞分裂。因此,研究CKS1家族基因参与甲壳动物性腺发育调节的分子机制具有重要的意义。从已构建的拟穴青蟹性腺EST数据库中筛选到CKS1B基因片段,继而克隆出其全长cDNA序列（Sp-CKS1B）,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,获得Sp-CKS1B的全长cDNA序列722 bp,其中5' UTR为82 bp,3' UTR为379 bp,开放阅读框261 bp,编码86个氨基酸,包含一段典型的CKS保守序列,属于CKS家族蛋白。在不同组织qRT-PCR结果显示,Sp-CKS1B 基因在O5期雌蟹卵巢中的表达水平最高,并与其他组织具有极显著差异（P<0.01）;同时,在性腺不同发育阶段的表达结果显示,Sp-CKS1B 基因在精巢T1期的表达量最高,其次为O5期,二者显著高于卵巢O1-O4期的表达水平（P<0.05）;而在精巢T3期的表达量最低,并显著低于T1、T2以及卵巢O4、O5期的表达水平（P<0.05）。结果说明了Sp-CKS1B在拟穴青蟹的性腺发育过程中可能具有十分重要的作用。

关键词: CDC28蛋白激酶调节亚基1B, 拟穴青蟹, 基因克隆, 组织表达, 性腺发育

Abstract: CDC28 protein kinase regulatory subunit 1B（CKS1B）is a member of the highly conserved cyclin kinase subunit 1（CKS1）protein family,and involves in cell division of eukaryotic organisms as a regulatory subunit of cyclin-dependent kinases. Therefore,it is of significance to study the function of CKS1 family genes involved in the molecular mechanism of regulating crustacean's gonad development. In the present study,the fragment of CKS1B gene was selected from the gonad EST library of S. paramamosain,and the full-length cDNA sequence of Sp-CKS1B was cloned,further the expressions of Sp-CKS1B in different tissues and different development stages of gonad were examined using qRT-PCR. As results,the full length cDNA of Sp-CKS1B was 722 bp,including a 5'untranslated region（5' UTR）of 82 bp,a 3'UTR of 379 bp,and an open reading frame of 261 bp encoding a protein of 86 amino acid. The protein contained a typical CKS conservative sequence belonging to the CKS family. Its expression level in the ovary of O5 stage female crabs was the highest,and presented extremely significant difference from other tissues（P<0.01）. Meanwhile,the expression level of Sp-CKS1B gene in T1 was the highest among the other stages of gonad development,followed by O5 stage,both of them were significantly higher than O1-O4 stage of female with significant difference（P<0.05）. Its lowest expression level was in the T3 stage,which was significantly lower than in T1,T2,O4,and O5 stages（P<0.05）. These results suggest thatSp-CKS1B may play a critical role in the gonad development of the mud crab.

Key words: CKS1B, Scylla paramamosain, gene cloning, tissue expression, gonad development

韩坤煌, 戴燕彬, 邹志华, 张子平, 王艺磊. CKS1B参与拟穴青蟹性腺发育的研究[J]. 生物技术通报, 2016, 32(11): 188-193.

HAN Kun-huang, DAI Yan-bin, ZOU Zhi-hua, ZHANG Zi-ping, WANG Yi-lei. On the Involvement of CKS1B in Gonad Development of Mud Crab Scylla paramamosain[J]. Biotechnology Bulletin, 2016, 32(11): 188-193.

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