Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand.
Aquatic Molecular Genetics and Biotechnology Research Team, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Khlong Nueng, Khlong Luang, Pathum Thani, 12120, Thailand.
Biochem Genet. 2021 Feb;59(1):114-133. doi: 10.1007/s10528-020-09993-8. Epub 2020 Aug 11.
The full-length cDNA of cyclin C of the giant tiger shrimp Penaeus monodon (PmCyC) was isolated by RACE-PCR. It was 1443 bp in length containing an open reading frame (ORF) of 804 bp and 267 deduced amino acids. Tissue distribution analysis indicated that PmCyC was more abundantly expressed in ovaries and testes than other tissues of female and male juveniles (P < 0.05). A pair of primers was designed, and an amplification product of 403 bp containing an intron of 123 bp was obtained. Polymorphism of amplified PmCyC gene segments of the 5th (3-month-old G5, N = 30) and 7th (5-month-old G7, N = 18) generations of domesticated juveniles was analyzed. Four conserved SNPs (T>C, T>C, G>A, and T>C) were found within the examined sequences. A TaqMan genotyping assay was developed for detection of a T>C SNP. Association analysis indicated that this SNP displayed significant association with body weight (P < 4.2e-10) and total length (P < 2e-09) of the examined G7 P. monodon (N = 419) with an allele substitution effect of 5.02 ± 0.78 g and 1.41 ± 0.19 cm, respectively. Juveniles with C/C (22.80 ± 2.51 g and 12.97 ± 0.53 cm, N = 19) and T/C (20.41 ± 0.93 g and 12.77 ± 0.21 cm, N = 129) genotypes exhibited a significantly greater average body weight and total length than those with a T/T genotype (14.72 ± 0.53 g and 11.37 ± 0.13 cm, N = 271) (P < 0.05).
通过 RACE-PCR 技术,从斑节对虾(Penaeus monodon)中克隆得到全长 cyclin C 基因(PmCyC)cDNA。该基因全长 1443bp,包含一个 804bp 的开放阅读框(ORF),可编码 267 个氨基酸。组织分布分析显示,该基因在雌、雄虾幼体的卵巢和精巢中表达量最高,显著高于其他组织(P < 0.05)。设计引物,成功扩增出一段 403bp 的基因片段,内含 123bp 的内含子。对 5 龄(3 月龄 G5,N = 30)和 7 龄(5 月龄 G7,N = 18)驯化幼虾的扩增产物进行多态性分析,共检测到 4 个 SNP(T>C、T>C、G>A 和 T>C)。开发 TaqMan 基因分型检测技术,对 T>C SNP 进行检测。关联分析表明,该 SNP 与 7 龄斑节对虾体重(P < 4.2e-10)和全长(P < 2e-09)显著相关,等位基因替换效应分别为 5.02 ± 0.78g 和 1.41 ± 0.19cm。与 T/T 基因型相比,C/C 基因型(22.80 ± 2.51g 和 12.97 ± 0.53cm,N = 19)和 T/C 基因型(20.41 ± 0.93g 和 12.77 ± 0.21cm,N = 129)个体的平均体重和全长显著增加(P < 0.05)。