梨形环棱螺qRT-PCR内参基因的筛选
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作者:
作者单位:

1.华中农业大学水产学院/农业农村部淡水生物繁育重点实验室,武汉 430070;2.柳州市渔业技术推广站,柳州 545006

作者简介:

郭雪朋,E-mail:1610351580@qq.com

通讯作者:

周小云,E-mail:zhouxy@mail.hzau.edu.cn

中图分类号:

S917.4

基金项目:

国家重点研发计划项目(2022YFD2400700)


Screening of optimal internal reference genes for qRT-PCR in Bellamya purificata
Author:
Affiliation:

1.College of Fisheries,Huazhong Agricultural University/Key Laboratory of Freshwater Animal Breeding,Ministry of Agriculture and Rural Affairs of the People’s Republic of China,Wuhan 430070,China;2.Liuzhou Aquaculture Technology Extending Station,Liuzhou 545006,China

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    摘要:

    为筛选梨形环棱螺(Bellamya purificata)在不同发育阶段、不同组织以及性别间各组织中表达稳定的最佳qRT-PCR内参基因,分别用geNorm、NormFinder、BestKeeper软件和比较ΔCt法,以及综合分析软件RefFinder评估actβ18Sgapdhef1αef1βrps3rpl4tubαtubβsdha共10个候选基因在梨形环棱螺的胚胎阶段、幼螺阶段和雌雄成螺各组织中的表达稳定性。结果显示,候选内参基因在胚胎和幼螺阶段的表达稳定性依次为gapdh>tubα>18S>ef1α>ef1β>rps3>actβ>tubβ>sdha>rpl4,在雌螺各组织中的表达稳定性依次为ef1β>tubα>rps3>ef1α>18S>actβ>gapdh>sdha>tubβ>rpl4,在雄螺各组织中的稳定性依次为ef1α>rps3>ef1β>rpl4>tubα>actβ>18S>gapdh>tubβ>sdha,在性别间各组织中的稳定性依次为ef1β>ef1α>rps3>tubα>18S>actβ>gapdh>rpl4>sdha>tubβ。上述结果表明,gapdhtubα为胚胎和幼螺阶段较适宜的qRT-PCR内参基因,ef1βtubα为雌螺组织间较适宜的qRT-PCR内参基因,ef1αrps3为雄螺组织间较适宜的qRT-PCR内参基因,ef1αef1β为性别间较适宜的qRT-PCR内参基因。

    Abstract:

    To select optimal reference genes to improve the accuracy of qRT-PCR analysis results in the freshwater snail Bellamya purificata,ten commonly used reference genes,including actβ18Sgapdhef1αef1βrps3rpl4tubαtubβ and sdha were chosen as candidate genes,and geNorm,NormFinder,BestKeeper,ΔCt and RefFinder were used to assess their expression stability during the embryonic and larval stages,as well as across different tissues and between sexes in adult individuals.The results showed that,the expression stability of the candidate genes during the embryonic and larval stages was ranked as follows:gapdh>tubα>18S>ef1α>ef1β>rps3>actβ>tubβ>sdha>rpl4.In terms of inter-tissue expression stability between female and male individuals,the expression stability of the candidate genes were ranked as ef1β>tubα>rps3>ef1α>18S>actβ>gapdh>sdha>tubβ>rpl4 for females,and ef1α>rps3>ef1β>rpl4>tubα>actβ>18S>gapdh>tubβ>sdha for males,respectively.In terms of tissue expression stability between sexes,the ranking was as follows:ef1β>ef1α>rps3>tubα>18S>actβ>gapdh>rpl4>sdha>tubβ.Consequently,gapdh and tubα are recommended as reference genes for qRT-PCR analysis during the embryonic and larval stages,ef1β and tubα are suggested as reference genes for qRT-PCR analysis across female snail tissues,ef1α and rps3 are suggested as reference genes for qRT-PCR analysis across male snail tissues,while ef1α and ef1β are the most appropriate for qRT-PCR analysis between sexes in B.purificata.

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郭雪朋,文衍红,罗福广,王志强,祝东梅,周小云.梨形环棱螺qRT-PCR内参基因的筛选[J].华中农业大学学报,2025,44(6):253-262

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  • 收稿日期:2025-01-17
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  • 在线发布日期: 2025-12-16
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