小鼠细小病毒非结构基因转基因小鼠的建立

TRANSGENIC MICE FROM FERTILIZED EGGS MICROINJECTED WITH PARVOVIRUS MINUTE VIRUS OF MICE NON-STRUCTURAL GENE

为探讨小鼠细小病毒（ＭＶＭ）非结构蛋白在ＭＶＭ感染中的抗肿瘤作用，酶切表达质粒ｐＵＬＢ３２３８获取该非结构基因，通过显微注射法接种入小鼠受精卵内制备转基因鼠。共注射受精卵７２０枚，选取存活受精卵２２５枚植入假孕小鼠输卵管，产仔１４只。转基因小鼠尾部组织ＰＣＲ法ＤＮＡ检测证明，其中４只整合靶基因。整合转基因的４只Ｇ０代小鼠与正常Ｃ５７ＢＬ／６Ｊ小鼠配种均可生产整合靶基因的小鼠。ＲＴ－ＰＣＲ法ｍＲＮＡ检测证明，Ｇ０Ａ６转基因鼠的肺、乳腺、颌下腺组织，Ｇ０Ａ１３转基因鼠的乳腺组织有靶基因转录。这一结果表明，ＭＶＭ非结构基因转基因小鼠模型已建立。

To investigate the role of parvovirus minute virus of mice (MVM) non-structural protein in the anti tumorigenesis of MVM infection, transgenic mice carrying MVM non-structural gene were produced by microinjecting the gene that contained the MMTV-LTR promoter and MVM non-structural gene. The microinjected fertilized eggs were 750, the choiced effective eggs were 225, and 14 transgenic mice were born. Transgenic mice were identified by subjecting tail DNA to amplification by PCR using a set of oligonucleotide as primers. RT-PCR analysis also affirmed the presence of transcript of the MVM non-structural gene in lung, mammary gland and salivary gland of G 0A6 and in mammary gland of G 013. These results demonstrate that transgenic mice model for MVM non-structural gene has been established.