VP_1intron敲除对pSVL WAP tPA暂时性乳腺定位表达的影响

Effect of knocking out VP_1 intron on brief specific expression of pSVL WAP tPA vector in mammary cells

在以ｃＤＮＡ表达外源蛋白时，表达载体中是否含有ｉｎｔｒｏｎ 会对表达造成一定的影响－ 利用基因工程手段将乳腺定位表达载体ｐＳＶＬＷＡＰｔＰＡ 中ＶＰ１ｉｎｔｒｏｎ 敲除，构建了表达载体ΔｐＳＶＬＷＡＰｔＰＡ－ 采用基因直接注射方法将两种表达载体分别注入家兔乳腺中，溶圈试验结果显示，ＶＰ１ｉｎｔｒｏｎ 去除后，ｔＰＡ 表达水平有所提高，分娩后第５ ｄ 乳中表达水平分别为４００ μｇ·Ｌ－１和６５０ μｇ·Ｌ－ １－

There are different effects of intron in vector on expression of foreign gene cDNA.On the basis of mammary gland specific expression vector pSVL WAP tPA,ΔpSVL WAP tPA vector was constructed by knocking out VP 1 intron.Two vectors were expressed in rabbit mammary gland by injecting vector into mammary.Milk obtained from rabbit contains biologically active t PA,and concentration were calculated to be about 400 μg·L -1 and 650 μg·L -1 on 5th day after childbirth,respectively.The result demonstrated that knocking out VP 1 intron can promote t PAcDNA expressing and the encoded protein can secrete into milk.