摘要
乳清酸蛋白 (WAP)和 β -酪蛋白 (β -Casein)是哺乳动物乳汁中的主要蛋白质 ,其基因 5′侧翼区常作为乳腺定位表达的调控序列。表达载体pSVL -WAP -tPA和pSVL - βb -tPA分别含 1.1kb的大鼠WAP启动子和 0 .6kb的牛β Casein启动子序列及t-PAcDNA。采用基因直接注射方法将 2种表达载体分别注入家兔乳腺中。溶圈试验结果显示 ,两种表达载体均能在兔乳腺细胞中得到表达 ,且以分娩后第 4天的乳汁中表达水平最高 ,分别为 45 0ng·mL-1和390ng·mL-1。为进一步研究t-PA基因乳腺定位表达调控和建立t-PA乳腺生物反应器奠定了基础。
Two mammary gland specific expression vectors were constructed with 1.1kb rat WAP or 0.6 kb bovine β Casein promoter and cDNA encoding human tissue type plasminogen activator(t PA).The vectors mixed with liposome were injected into the mammary glands of pregnant rabbits through the mammary gland duct and the rabbits were milked at day 1,2,4,6,8 and 10 after parturition.Fibrin clot lysis assay showed that the promoter of WAP and β Casein were able to direct low level expression of t PA cDNA.The expression level was 450ng·mL 1 and 390ng·mL 1 on day 4?respectively,reaching the higest level.It provides useful information for studying the regulation of specific expression in mammary gland and the establishment of t PA mammary gland bioreactor. [WT5HZ]
出处
《洛阳农业高等专科学校学报》
2001年第2期79-81,共3页
Journal of Luoyang Agricultural College
基金
国家"86 3"计划资助项目部分内容 (Z2 1- 0 4- 0 3)