斑马鱼MC1R基因克隆鉴定、生物信息学及序列变异分析

打开文本图片集

中图分类号：S965.819 文献标志码：A

Clong，biomatics sequence variation analysis of MCiR geneDaniorerio

（Colege ofAgroesryEngeerg Planng， FanjgMounta，， ，Cha）

Abstract：【Objective】This study aimed to clone identify codg sequence（CDS）of melanocort-1receptor（MC1R）gene zebrafish（Danio rerio），toconduct biomatics sequence variation analysis，soas to provide fundamental materials elucidatg biological function of MC1R gene regulation mechanismsof zebrafish bodycolor traits.【Method】Usg muscle tissueDNA from 219 samples ofred，yellow， blue zebrafish populations as templates，CDS sequences of MC1R gene of zebrafish were obtaed through PCR amplification，clong，sequencg，asembly to perm biomaticsanalysis gene sequencevariationanalysis.【ResultThe fulllength ofMCiR genewas 1404bp ，witha CDS of972bp，encodg 323amo acid residues.TheMC1R prote contaed seven transmembrane domas，four extracellarregions， four tracelularregions，exhibitg atypial cel surface receptor-like structure.The prote was maly localized to plasma membrane possssd four potential Nglycosylationsites26phosphorylationsites.The zebrafish MC1R gene showed high similaritytothatofMegalobrama amblycephalaAnabarilius grahami，demonstratg nearestgeneticdistanceclosegeneticrelationships.Two variant sites were identified red，yellow， blue zebrafish populations：one variant site （ ）waslocated CDS exhibited bothTTTC genotypes，without causg anychange amo acid； or variant site was located 3'-untranslated region（3'-UTR）（ g.276A>T ）exhibitedbothAAAT genotypes.Thefrequenciesof two variants， g.334T>C g.276A>T ，were consistent across color groups.【Conclusion】The MC1R gene sequence is conserved somewhat diverse across fish species，suggestg that prote structure isrelatively conserved functionallysimilar acrosspecies.The MClR gene can be used as an effective molecular marker studyg evolutionaryrelationshipsofdiferentfishsystems.Itisrelativelyconserved red，yellow， bluezebrafish populations， contas two variant sites（ g.276A>T g.334T>C ）which maybe lkage.

words：Danio rerio；melanocort-1 receptor gene（MC1R）；variant sites；body color

Foundation items：National Natural Science Foundation of Cha（32060274）； Science Technology Plan Project of Provce（QKHJC-ZK[2022]Yiban559）； High-level Innovative Talent（Thous Level）Project （2024-[2022]-048）；Innovation Entrepreneurship Trag Program Colege Students （S2024106 651872）

0 引言

【研究意义】斑马鱼（Daniorerio）隶属于硬骨鱼纲（Osteichthyes）鲤形目（Cyprimes）鲤科（Cypridae）短担尼鱼属（Danio）（PattersonParichy，2019），其体色表型丰富且易于繁殖，不仅具有较高的观赏价值，也是研究鱼类体色遗传机制的重要模式生物（Irionetal.，2016，杨倩等，2023）。（剩余19530字）