摘要
目的 制备乳腺生物反应器所必需的乳腺特异性表达的调控序列 ,并验证其指导外源基因表达的能力。方法 用PCR法从奶牛染色体上分 5段扩增出了全长 8 2Kb的牛 β 乳球蛋白基因 ,包括 1 8Kb的 5′侧翼区、1 7Kb的 3′侧翼区及 4 7Kb的gDNA区。扩增出的各片段克隆到T 载体上 ,酶切鉴定及序列分析均证实了所扩增片段的正确性。将五个片段与荧光素酶cDNA拼接成荧光素酶瞬时表达载体并在小鼠乳腺中瞬时表达。结果 注射荧光素酶瞬时表达载体的小鼠乳汁中明显测出了荧光素酶活性。结论 所克隆的牛 β 乳球蛋白基因表达调控序列能够指导外源基因在小鼠乳腺中表达。
Objective\ To get the regulatory elements which are essential for generating mammary gland bioreactors,and identify their effect for regulating the expression of foreign gene.Methods\ The whole 8.2 Kb bovine beta-lactoglobulin gene was amplified by PCR method,including the 1.8 Kb 5′ flanking region,4.7 Kb gDNA region,1.7 Kb 3′ flanking region.All the amplified fragments were cloned in T-vectors and were proved to be correct by restrction enzyme digestion and sequencing analysis.The five fragments and the luciferase cDNA were ligated together to get the luciferase temporary expression vector,which was temporarily expressed in the mammary gland of mice.Results\ The luciferase activity was detected in the milk of mice whose mammary glands were injected with the expression vector.Conclusion\ The cloned bovine BLG gene could direct foreign gene expression in the mammary gland of mice.\;
出处
《中国实验动物学报》
CAS
CSCD
2001年第2期78-82,共5页
Acta Laboratorium Animalis Scientia Sinica
基金
国家863计划! (Z2 1 0 3 0 1 )
