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构建了包含山羊β-酪蛋白基因启动区5′端上游调控序列和外显子1, 2及内含子1在内的乳腺表达载体, 并与人血清白蛋白(hALB)小基因(含全长cDNA序列及其内含子1)构建成融合基因. 鼠尾静脉注射实验表明, 该融合基因能在小鼠乳腺中特异表达. 应用显微注射方法将该融合基因导入小鼠受精卵, 并将受精卵移植于假孕母鼠, 共获得33只F0代小鼠, 经PCR和Southern印迹杂交证实, 有8只小鼠(5♀, 3 ♂ )基因组中整合有hALB基因, 整合率为24.2%(8/33). Western blot分析表明, 在5只雌性整合小鼠中有3只表达了hALB蛋白, 放射免疫法测定其乳汁中hALB含量分别为3.54, 0.21和3.03 g/L.
A mammary gland expression vector containing the 5 ’upstream regulatory sequence of the promoter region of goat β-casein gene and exons 1, 2 and intron 1 was constructed and compared with the human serum albumin (hALB) minigene The long cDNA sequence and its intron 1) were constructed into fusion gene.The mouse tail vein injection experiments showed that the fusion gene can be expressed specifically in the mouse mammary gland.The microinjection method was used to transfer the fusion gene into the mouse zygotes A total of 33 F0 mice were obtained by transplanting fertilized eggs into fake pregnant mice. Confirmation of PCR and Southern blot hybridization showed that hALB gene was integrated into the genome of 8 mice (5 ♀, 3 ♂) with an integration rate of 24.2 % (8/33) .Western blot analysis showed that 3 out of 5 female mice expressed hALB protein, and the hALB content in breast milk was 3.54, 0.21 and 3.03 g / L respectively by radioimmunoassay.