转绿色荧光蛋白标记基因猪整合检测的研究

目的]为转基因猪生产的整合效率、外源目的基因表达水平、目的基因在猪体内组织特异性表达分布等相关领域的进一步研究奠定基础。方法]用显微注射法将GFP标记的转基因猪表达载体PM1导入湖北白猪和通湖猪1-细胞、2-细胞胚胎中,胚胎移植妊娠后生产仔猪,取耳或尾组织样提取基因组总DNA,用PCR法检测PM1在猪基因组中的整合情况。结果]在所有生产的78头仔猪中,有6头猪用PCR检测为转PM1基因阳性猪,这6头猪在3次PCR扩增反应中都得到650 bp目的扩增片段。其他样品没有得到目的扩增片段的为阴性猪。结论]载体PM1可以用于携带外源基因进行转基因猪生产。

Study on Integration Detection of Transgenic Pig Marked with Green Fluorescent Protein

Objective] The aim was to lay the foundation for further study on the related fields such as integration efficiency of transgenic pig production,expressive level of foreign target gene,specific expressive distribution of target gene in the tissue of pig and so on.Method] The transgenic pig expression vector PM1 marked with GFP gene was introduced into the 1-cell or 2-cell embryos of Hubei White pigs and Tonghu pigs by microinjection to produce piglets by embryonic implantation.The total DNA from genome was extracted from the pig ears and tails and the integration situation of PM1 in the pig genome was detected by PCR. Result] Among the 78 produced piglets,there were 6 transgenic positive pigs detected by PCR and the amplified target fragment of 650 bp from the 6 pigs was all obtained by 3 PCR amplified reactions.Other samples which did not obtain the amplified target fragment were negative pigs.Conclusion] The PM1 vector could be used to carry foreign genes to produce the transgenic pigs.