农业生物技术学报2025，Vol.33Issue(12)：2668-2676,9.DOI:10.3969/j.issn.1674-7968.2025.12.010
瓯江彩鲤OCA2基因的克隆与初步功能研究
Cloning and Preliminary Functional Study of the OCA2 Gene in Oujiang Color Common Carp(Cyprinus carpio var.color)
摘要
Abstract
The OCA2(oculocutaneous albinism Ⅱ)gene is an important gene in melanin biosynthesis,and involved in pigmentation in the eye,skin,and hair colors of animals.Mutations in this gene could result in oculocutaneous albinism type Ⅱ in human.The black patchs on the body surface are key phenotypic characteristics in Oujiang color common carp(Cyprinus carpio var.color),however,the genetic basis and underlying mechanisms of their formation remain unclear.In this study,The high-generation selected strains of Oujiang color common carp was used to perform the cloning,expression and promoter transcriptional activity studies of the OCA2 gene,and gene knockout experiment was carried out by the CRISPR/Cas9 technology.The results revealed that the OCA2 gene consisted of 23 exons and 22 introns,encoding 834 amino acids with over 97%homology with the other fish species in Cyprinid family,which indicated highly structural and functional conservation.The qPCR analysis of the OCA2 gene showed that its expression levels were significantly higher in the eye and skin tissues in the whole white with black patch than those in the whole white color types in Oujiang color common carp.The dual-luciferase reporter assays indicated a complex regulatory mechanism in the promoter region of the OCA2 gene,and a key regulatory element during the 600～1 200 bp fragment before the start codon.CRISPR/Cas9-mediated knockout of exon and promoter regions in the OCA2 gene resulted in the numbers reduction by 70%and the areas decreasing by 40%of melanocytes from surface skin and eye tissues in the mutant.This study revealed that the OCA2 gene plays an important role in pigmentation in Oujiang color common carp.关键词
瓯江彩鲤/OCA2基因/体色/黑色素Key words
Oujiang color common carp/OCA2(oculocutaneous albinism Ⅱ)gene/Body color/Melanin分类
农业科技引用本文复制引用
曾春晓,柯景,张鹏辉,陈慧帆,陈晓雯,王军,王成辉..瓯江彩鲤OCA2基因的克隆与初步功能研究[J].农业生物技术学报,2025,33(12):2668-2676,9.基金项目
淡水鱼种质资源生态化保存与更新(2022YFD2400102) （2022YFD2400102）
