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分别将构建的人顶体蛋白酶原基因 (humanproacrosin ,hACR) 5’侧翼序列 ,第 1外显子和编码 β 半乳糖苷酶的基因 (lacZ)的融合基因 (载体 1 ) ,以及hACR基因 5’侧翼序列与报告基因lacZ的融合基因 (载体 2 ) ,通过显微注射受精卵法制备转基因小鼠 ,结果如下 :载体 1 :注射 89只卵 ,移植入 9只母鼠单侧输卵管中 ,3只怀孕 ,产仔9只 ,存活 7只 ,基因组Southern杂交检出整合有外源基因的阳性小鼠 1只 ,阳性率 1 4 .3% ;子 1代 1 5只 ,基因组Southern杂交 5只阳性。载体 2 :注射 2 35只卵 ,移植入 1 4只母鼠单侧输卵管中 ,4只怀孕 ,产仔 1 1只 ,存活 1 0只 ,基因组Southern杂交检出整合有外源基因的小鼠 1只 ,阳性率 1 0 .0 %。子 1代小鼠 1 0只 ,基因组Southern杂交 3只阳性。进一步将检测lacZ在转基因小鼠体内的表达。
The 5 ’flanking sequence of human acrosin gene (hACR), the fusion gene (vector 1) of exon 1 and the gene encoding β-galactosidase (lacZ), and the 5’ The flanking sequence and the reporter gene lacZ fusion gene (vector 2), transgenic mice were prepared by microinjection of fertilized egg method, the results are as follows: Vector 1: 89 eggs were injected, transplanted into nine unilateral fallopian tubes, 3 Nine pregnant and 18 offspring survived, and 1 positive mouse with positive foreign gene was detected by Southern hybridization. The positive rate was 14.3%, 15 in the first generation and 5 in the Southern hybrid. Vector 2: 2 35 eggs were injected and transplanted into unilateral fallopian tubes of 14 maternal mice, 4 pregnant, 11 litters, and 10 surviving mice. Genomic Southern blotting confirmed that mice with integrated exogenous gene 1 Only, the positive rate of 1.0%. 10 mice in the first generation and three in the genomic Southern hybrid. Further examination of lacZ expression in transgenic mice will be performed.