CAJ | 학술논문

该研究以斑节对虾（ Penaeus monodon）cDNA文库中的CDK2片段为基础，利用RACE技术获得PmCDK2的cDNA全长，并利用生物信息学对其进行了分析。PmCDK2基因全长1679 bp，包括编码306个氨基酸的921 bp的开放阅读框（ ORF），258 bp的5＇UTR和500 bp的3＇UTR。同源性分析显示，PmCDK2与罗氏沼虾（ Macrobrachi-um rosenbergii）等物种的CDK2具有较高的同源性。使用荧光定量PCR技术研究了PmCDK2的mRNA在不同组织和卵巢发育各时期的相对表达量和变化模式，结果显示，PmCDK2在斑节对虾的脑、心、淋巴、肝胰腺、性腺等组织中均有表达，其中精巢中表达量显著高于其他组织；卵巢发育阶段表达分析则显示PmCDK2在Ⅲ期表达量最高，其次是Ⅳ期。通过原核表达技术成功获得了含有6个His标签的融合蛋白，并对融合蛋白进行 Western Blot试验，初步探讨了重组蛋白表达情况。结果表明，PmCDK2基因在斑节对虾卵巢发育过程中可能发挥了重要作用，该结果对进一步探究斑节对虾卵巢发育机理提供了一定的理论依据。
해연구이반절대하（ Penaeus monodon）cDNA문고중적CDK2편단위기출，이용RACE기술획득PmCDK2적cDNA전장，병이용생물신식학대기진행료분석。PmCDK2기인전장1679 bp，포괄편마306개안기산적921 bp적개방열독광（ ORF），258 bp적5＇UTR화500 bp적3＇UTR。동원성분석현시，PmCDK2여라씨소하（ Macrobrachi-um rosenbergii）등물충적CDK2구유교고적동원성。사용형광정량PCR기술연구료PmCDK2적mRNA재불동조직화란소발육각시기적상대표체량화변화모식，결과현시，PmCDK2재반절대하적뇌、심、림파、간이선、성선등조직중균유표체，기중정소중표체량현저고우기타조직；란소발육계단표체분석칙현시PmCDK2재Ⅲ기표체량최고，기차시Ⅳ기。통과원핵표체기술성공획득료함유6개His표첨적융합단백，병대융합단백진행 Western Blot시험，초보탐토료중조단백표체정황。결과표명，PmCDK2기인재반절대하란소발육과정중가능발휘료중요작용，해결과대진일보탐구반절대하란소발육궤리제공료일정적이론의거。
The full-length cDNA sequence of CDK2 from black tiger shrimps(Penaeus monodon)(denoted as PmCDK2)was obtained by high throughput transcriptome sequencing and RACE. The cDNA of PmCDK2 was 1 679 bp,including an open reading frame( ORF) of 921 bp encoding a polypeptide of 306 amino acids,a 5'UTR of 258 bp and a 3'UTR of 500 bp. Blast and phylogenetic analyses suggest that PmCDK2 is a new member of the shrimp CDKs family. PmCDK2 mRNA expression was detected in seven tissues by real-time PCR. Expression was the highest in the testis and moderate in the ovary,lymph,muscle and so on. While at different stages of ovary develop-ment,PmCDK2 expressed most at StageⅢ,indicating that PmCDK2 might play an important role in ovary development for black tiger shrimp. The results provide insight into the function of PmCDK2 in regulation of ovarian development of black tiger shrimp.