Qiu Lihua, Jiang Shigui, Zhou Falin, Huang Jianhua, Guo Yihui
Biotechnology and aquiculture Laboratory, The South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 231 Xingangxi Road, Guangzhou, 510300, P.R. China.
Mol Biol Rep. 2023 Oct;50(Suppl 1):S1-S8. doi: 10.1007/s11033-006-9052-4. Epub 2007 Jan 24.
The techniques of homology cloning and anchored PCR were used to clone the cyclin B gene from black tiger shrimp. The full length cDNA of black tiger shrimp cyclin B (btscyclin B) contained a 5' untranslated region (UTR) of 102 bp, an ORF of 1,206 bp encoding a polypeptide of 401 amino acids with an estimated molecular mass of 45 kDa and a 3' UTR of 396 bp. The searches for protein sequence similarities with BLAST analysis indicated that the deduced amino acid sequence of btscyclin B was homological to the cyclin B of other species and even the mammalians. Two conserved signature sequences of cyclin B gene family were found in the btscyclin B deduced amino acid sequence. The temporal expressions of cyclin B gene in the different tissues, including liver, ovary, muscle, brain stomach, heart and intestine, were measured by RT-PCR. mRNA expression of cyclin B could be detected in liver, ovary, muscle, brain, stomach, heart and strongest in the ovary, but almost not be detected in the intestine. In ovarian maturation stages, the expression of btscyclin B was different. The result indicated that btscyclin B was constitutive expressed and played an important role in the cell division stage.
采用同源克隆和锚定PCR技术从黑虎虾中克隆细胞周期蛋白B基因。黑虎虾细胞周期蛋白B(btscyclin B)的全长cDNA包含一个102bp的5'非翻译区(UTR)、一个1206bp的开放阅读框(ORF),编码一个401个氨基酸的多肽,估计分子量为45kDa,以及一个396bp的3'UTR。通过BLAST分析搜索蛋白质序列相似性表明,btscyclin B推导的氨基酸序列与其他物种甚至哺乳动物的细胞周期蛋白B同源。在btscyclin B推导的氨基酸序列中发现了细胞周期蛋白B基因家族的两个保守特征序列。通过RT-PCR检测细胞周期蛋白B基因在肝脏、卵巢、肌肉、脑、胃、心脏和肠道等不同组织中的表达时间。在肝脏、卵巢、肌肉、脑、胃、心脏中均可检测到细胞周期蛋白B的mRNA表达,在卵巢中表达最强,但在肠道中几乎检测不到。在卵巢成熟阶段,btscyclin B的表达有所不同。结果表明,btscyclin B组成性表达,在细胞分裂阶段发挥重要作用。