| 摘要: |
| 由牛β-Casein启动子与大鼠WAP启动子融合构建了t-PA cDNA乳腺定位表达载体pSVL-β△WAP-PA,以研究t-PA乳腺定位表达的调控。结果表明，将表达载体pSVL-βb-PA,pSVL-WAP-PA及pSVL-β△WAP-PA分别直接注入怀孕的家兔乳腺管中，溶圈试验显示3种表达载体均能在家兔乳腺中表达，且以分娩后第4天的乳汁中表达水平最高，分别为390，430和670 ng/mL。证明0.6 kb的牛β-Casein上游调控序列具有一定的定位表达调控能力。融合启动子较单纯的β-Casein或WAP启动子调控下的t-PA cDNA表达水平高。 |
| 关键词: β-Casein基因 乳清酸蛋白基因；t-PA cDNA；乳腺定位表达 |
| DOI： |
| 分类号: |
| 基金项目:国家“863”计划资助项目(Z21-04-03) |
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| Fused promoter regulating t-PA cDNA expression in mammary gland |
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| Abstract: |
| In order to study the expressing regulation of t-PA cDNA in mammary gland,two specific expression plasmids,pSVL-βb-PA and pSVL-β△WAP-PA,containing t-PA cDNA gene,were constructed by fusing with bovine β-Casein promoter,respectively.The three plasmids,pSVL-βb-PA,pSVL-WAP-PA and pSVL-β△WAP-PA were mixed with liposome and injected directly into pregnatn rabbit mammary gland duct.Fibrin clot lysis assay showed that the three plasmids could express t-PA in rabbit milk.The expression level was 390,430 and 670 ng/mL on day 4,respectively.It occured that 0.6 kb bovine 5’regulatory region of β-Casein gene was able to direct low level expression of t-PA cDNA in mammary gland,and the expression level of t-PA cDNA regulated by fused promoter was higher than β-Casein or WAP promoter.It also provides the useful information for studing the regulation of specific expression in mammary gland and the extablishment of t-PA mammary gland bioreactor. |
| Key words: Casein gene WAP gene t-PA cDNA mammary expression |
