黑龙江动物繁殖2025，Vol.33Issue(4)：23-28,6.DOI:10.19848/j.cnki.ISSN1005-2739.2025.08.0004
ERV1-2B_SSc-LTR-EGFP报告系统的建立及猪克隆胚胎ERVs转录活性的监测
Establishment of ERV1-2B_SSc-LTR-EGFP reporter system to monitor PERV transcriptional activity in cloned porcine embryos
摘要
Abstract
Somatic cell nuclear transfer(SCNT)technology is the core technique for producing transgenic pig models,however,the developmental capacity of cloned porcine embryos by SCNT is relatively limited,suggesting the significance of further investigation on the expression patterns of endogenous retroviruses(ERVs)in early embryos.This study aimed at constructing a reporter system using the porcine genomic endogenous retrovirus regulatory element ERV1-2B_SSc-LTR to drive EGFP expression.Positive fibroblast cell lines were obtained through lipofection,cloned embryos were constructed by SCNT,with real-time monitoring of EGFP expression under fluorescence microscopy.Results showed that the ERV1-2B_SSc-LTR-EGFP reporter system was successfully constructed;EGFP expression was observed in 48%of cloned embryos at the 4-cell stage,demonstrating the promoter activity of the EGFP;The blastocyst formation rate and cell number in EGFP-positive embryos were significantly higher than those in EGFP-negative embryos[(19.5±1.7)%vs.(14.8±0.5)%,(40.0±2.1)cells vs.(30.0±1.9)cells,P<0.05],respectively.These findings suggest that this reporter system can be used to dynamically monitor ERV activity and assist in evaluating the quality of SCNT embryos.关键词
体细胞核移植/长末端重复序列/绿色荧光蛋白报告系统/克隆/胚胎Key words
somatic cell nuclear transfer/long terminal repeat/EGFP reporter system/clone/embryos分类
生物科学引用本文复制引用
付博,马红,汪亮,郭镇华,王芳,刘娣..ERV1-2B_SSc-LTR-EGFP报告系统的建立及猪克隆胚胎ERVs转录活性的监测[J].黑龙江动物繁殖,2025,33(4):23-28,6.基金项目
黑龙江省省属科研院所科研业务费项目(CZKYF2024-1-B006) （CZKYF2024-1-B006）
