| 摘要: |
| 通过同源克隆以及5′/3′RACE 的方法扩增得到大菱鲆(Scophthalmus maximus)细胞周期蛋白依赖激酶基因cdk1和cdk6的全长cDNA序列, 在mRNA水平上分析了它们组织表达特征, 并对比分析了静水压处理对其胚胎期表达的影响。结果显示, cdk1基因全长cDNA序列为1281bp(GenBank登录号:KP339306), 编码区为912bp; cdk6基因cDNA全长1400bp(GenBank 登录号: KT186374), 编码区长度为978bp。组织RT-PCR分析表明, cdk1基因表达具有广泛性, 在性腺、肾脏等生长发育较快的组织中表达量较高; cdk6基因也在多个组织中表达, 在性腺中表达量最高。通过Real time RT-PCR检测基因在胚胎中的表达水平发现, 静水压处理组cdk1、cdk6在胚胎发育中, 总体变化趋势与对照组类似。大菱鲆胚胎对照组中cdk1在原肠期以前相对神经胚期及以后时期表达量较高, 静水压处理组基因表达变化与对照组趋势相同; 对照组中cdk6在原肠期出现表达高峰, 但静水压处理组在原肠期表达量相对较低。静水压处理对cdk1和cdk6 的表达量的影响不同, 这可能与基因还有除调控细胞周期的其他功能有关。为解析这两个基因在大菱鲆胚胎发育中的作用及其机制提供了参考。 |
| 关键词: 大菱鲆(Scophthalmus maximus) cdk1和cdk6 克隆 组织 胚胎 静水压 RT-PCR |
| DOI：10.11759/hykx 20150726003 |
| 分类号: |
| 基金项目:国家863 计划资助项目(2012AA10A402); 山东省自然科学基金资助项目(ZR2011CM035); 国家自然科学基金资助项目(31502156); 山东省科技发展计划资助项目(2013GHY11526); 鲆鲽类产业技术体系课题(NYCYTX-50-G03) |
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| Cloning and expression analysis of cdk1 and cdk6 in Scophthalmus maximus |
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GUO Jia-wei,ZHU Xiang-ping,WU Zhi-hao,SONG Zong-cheng,FAN Zhao-fei,TAN Xun-gang,YOU Feng
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| Abstract: |
| Whole cDNA sequences of cyclin-dependent protein kinase genes (cdk1 and cdk6) in turbot (Scophthalmus maximus) were sequenced using homologous cloning and 5′/3′-RACE. The open reading frames of cdk1 and cdk6 were 912 bp (GenBank accession number, KP339306) and 978 bp (GenBank accession number, KT186374), respectively. Gene expression in turbot tissues was analyzed using RT-PCR. The results showed that both cdk1 and cdk6 were widely expressed in several turbot tissues. cdk1 expression levels were higher in the gonads and kidneys than in other organs, and cdk6 expression level was the highest in the gonads. cdk1 and cdk6 expression levels in turbot embryos were studied using real-time RT-PCR. In the control group, cdk1 expression levels were higher at the gastrula stage and earlier compared to those at the neurula stage and later, and cdk6 expression level at the gastrula stage was the highest. Trend variation pertaining to cdk1 and cdk6 expression levels in the hydrostatic pressure treatment group (tetraploid induction group) was similar to that in the control group, except for the expression level, which was lower in the treatment group than in the control group. Hydrostatic pressure treatments had different effects on cdk1 and cdk6 expression levels, suggesting cdk1 and cdk6 in turbot to have different functions. The above results could provide a basis for the analysis of functions and mechanisms of the two genes in turbot embryo development. |
| Key words: Scophthalmus maximus cdk1 and cdk6 cloning tissue embryo hydrostatic pressure treatment RT-PCR |
