NRAMP1基因过表达山羊的生产及血液生理生化和生长性能测定  

Production and Determination of Blood Physiology,Biochemistry and Growth Performance of Goats(Capra hircus)Overexpressing the NRAMP1 Gene

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作  者:王聪亮 陈文博 万仕成 李剑南 任赵飞 韩斌 李娜 雷安民 杜晓敏 屈雷[2] 朱海鲸[2] 华进联 WANG Cong-Liang;CHEN Wen-Bo;WAN Shi-Cheng;LI Jian-Nan;REN Zhao-Fei;HAN Bin;LI Na;LEI An-Min;DU Xiao-Min;QU Lei;ZHU Hai-Jing;HUA Jin-Lian(Shaanxi Stem Cell Engineering and Technology Research Center,College of Veterinary Medicine,Northwest A&F University,Yangling 712100,China;Shaanxi Province Engineering&Technology Research Center of Cashmere Goats,Yulin University,Yulin 719000,China;Yulin Animal Husbandry and Veterinary Service Center,Yulin 719000,China)

机构地区:[1]西北农林科技大学动物医学院陕西省干细胞工程技术研究中心,杨凌712000 [2]榆林学院陕西省绒山羊工程技术研究中心,榆林719000 [3]榆林市畜牧兽医服务中心,榆林719000

出  处:《农业生物技术学报》2025年第4期837-846,共10页Journal of Agricultural Biotechnology

基  金:国家自然科学基金(32072806,32372970);国家重点研发计划(2022YFD1302201);陕西省畜牧科技示范和畜牧专项(20221086,20230978);陕西省教育厅重大项目(22JY075);陕西省农业攻关核心技术(2024NYGG005-6)。

摘  要:天然抗性相关性巨噬细胞蛋白1(natural resistance-associated macrophage protein 1,NRAMP1)与结核病、布鲁氏菌病等多种胞内菌感染抗性相关。本研究旨在制备NRAMP1基因过表达山羊(Capra hircus),并对基因过表达山羊的血液生理生化和生长发育指标进行比较分析,为抗布鲁氏菌病模型建立和转基因动物遗传育种提供研究模型与平台。基于本实验室前期构建的山羊NRAMP1基因慢病毒表达载体,利用山羊超数排卵和原核胚胎显微注射技术进行NRAMP1基因过表达山羊的生产,监测评估NRAMP1基因过表达对山羊血液生理生化变化和生长发育的影响。结果显示,成年陕北白绒山羊以210IU促卵泡激素(follicle-stimulating hormone,FSH)剂量进行超数排卵的原核胚胎回收率显著高于270 IU(P<0.05);原核胚胎显微注射后移植共获得15只山羊,经PCR鉴定,3只为阳性基因过表达山羊;qRTPCR和Western blot鉴定结果表明,NRAMP1在基因过表达山羊睾丸组织中的mRNA和蛋白表达水平显著增加(P<0.05);血液生理与生化测定结果表明,NRAMP1基因过表达山羊与野生型山羊的各项血液生理与生化指标差异不显著(P>0.05);生长性能测定结果表明,NRAMP1基因过表达山羊与野生型山羊初生重、1~6月龄体重和3~6月龄体高、体长等生长性状差异不显著(P>0.05)。本研究成功构建NRAMP1基因过表达山羊模型,且NRAMP1基因过表达不影响山羊自身健康。本研究为培育抗布鲁氏菌病山羊及其抗病机制研究提供了科学依据。Natural resistance-associated macrophage protein 1(NRAMP1)has been demonstrated to be linked with resistance to various intracellular mycobacterial diseases,including tuberculosis and brucellosis.This study aimed to develop goats(Capra hircus)with NRAMP1 gene overexpression and to conduct a comparative analysis of their blood physiology,biochemistry,and growth and development indices,which provides a research model and platform for establishing a brucellosis resistance model and for the genetic breeding of transgenic animals.Based on the lentiviral vector constructed in our laboratory for goat NRAMP1gene,NRAMP1 gene overexpression goats were produced using the techniques of supernumerary ovulation and prokaryotic embryo microinjection,and subsequently monitored and evaluated the effects of this overexpression on physiological and biochemical changes in the blood,as well as on the growth and development of the goats.The results showed that the recovery rate of prokaryotic embryos was significantly higher than that observed at a dose of 270 IU for supernumerary ovulation when administering 210 IU folliclestimulating hormone(FSH)in adult Shaanbei white cashmere goats(P<0.05).A total of 15 goats were obtained through the transplantation of prokaryotic embryos following microinjection,3 of which were identified as positive gene overexpression goats via PCR,and then q RT-PCR and Western blot analyses revealed that NRAMP1 expression was significantly elevated at both the mRNA and protein levels in the testicular tissues of the NRAMP1 overexpression goats(P<0.05).The results of blood physiological and biochemical measurements indicated that there were no significant differences in blood physiological and biochemical indices between NRAMP1 overexpressing goats and wild-type goats(P>0.05).Similarly,the results of growth performance measurements revealed that the differences in growth traits,including newborn weight,body weight from 1~6 months of age,and body height and length from 3~6 months of age,between NRAMP1 gene over

关 键 词:山羊 天然抗性相关性巨噬细胞蛋白1(NRAMP1) 过表达 慢病毒 显微注射 布鲁氏菌病 

分 类 号:S188[农业科学—农业基础科学]

 

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