中文摘要：

本研究旨在探讨内源性A型精原干细胞介导法制备转抗黏液病毒基因A（Myxovirus resistance gene,MxA）基因小鼠的可行性。将pcDNA-MxA质粒和新型转染试剂ExGen500混悬后分5点注射至7日龄的ICR公鼠两侧睾丸组织内,睾丸内基因注射（testis gene transfer,TGT）鼠在性成熟后的不同阶段（6、12及24周龄）与正常母鼠交配,检测后代外源基因整合及表达情况,选取表达MxA基因的小鼠进行H5N1亚型禽流感病毒攻毒试验,观察小鼠的健康状况并观察肺及气管的组织病变情况。结果表明,TGT鼠和正常母鼠交配后能稳定产生转基因后代小鼠,2只TGT鼠在6、12及24周龄交配后代整合率分别为11.11%（2/18）、11.76%（2/17）、11.54%（3/26）及13.64%（3/22）、13.33%（2/15）、11.76%（2/17）。2只TGT鼠后代外源基因平均整合率分别为11.47%及12.91%,之间差异不显著（P〉0.05）。RT-PCR检测表明,MxA基因整合小鼠中有21.43%（3/22）测到MxA的表达;所制备的MxA表达鼠H5N1亚型禽流感病毒攻毒后全身症状、肺及气管病变程度明显比正常鼠轻。试验结果表明体内精原干细胞介导的转基因方法是一种可行的、具有很大应用前景的转基因方法,制备的转MxA基因小鼠对H5N1亚型禽流感病毒具有一定的抵抗力。

英文摘要：

The aims of this study were to explore the feasibility and stability of producing MxA transgenic mice mediated by Type-A spermatogonia in vivo.The pcDNA-MxA plasmid and transfection reagent ExGen500were suspended and injected into testicle tissue of 7-day-old male ICR mice from 5different points;the testis-mediated gene transfer（TGT）mice mated with wild-type female mice at different sexual maturity stages（6-,12-and 24-week-old）,to detect the integration and expression of foreign gene in offspring.The mice expressed MxA were chosen for challenge test,health status and respiratory system tissue pathological changes were observed.Detection result of MxA integration by PCR and Southern blot in the F1-generation mice showed that foreign gene integration rates of 2TGT mice at different sexual maturity stages（6-,12-,and 24-week-old）were 11.11%（2/18）,11.76%（2/17）,11.54%（3/26）and 13.64%（3/22）,13.33% （2/15）,11.76%（2/17）,respectively.The average integration rates in two groups were 11.47% and 12.91%,the difference was not significant（P〉0.05）.RT-PCR detection result showed that MxAgene was expressed in 3of 14integration mice.Infection challenge test with H5N1influenza strains results showed that constitutional symptom and pathologic change degree of transgenic mice were more slighter than that of wild-type mice.The results indicated that the method of type-A spermatogonia mediated gene transfer shows bright future for application because it is a feasible and reproducible transgenic approach.MxAtransgenic mice have disease resistance to H5N1subtype avian influenza viruses to some degree.