泌乳乳腺作为真核基因的有效表达系统研究

周雪艳; 张守峰; 姚伟; 任文陟; 张菲; 扈荣良

doi:10.11847/zgggws2006-22-03-40

泌乳乳腺作为真核基因的有效表达系统研究

Study on lactating mammary gland as an efficient eukaryotic gene expression system

摘要

摘要:
目的探讨泌乳腺组织作为真核基因快速表达系统的可行性。
方法以蚓激酶(LK)作为目的标志基因, 分别构建了在泛组织嗜性的巨细胞病毒(CMV)立即早期启动子、逆转录病毒长末端重复序列(LTR)和组织特异性的beta-酪蛋白启动子调控下的3种真核表达载体pICLK、pLLKSN和pIbLK。制备各重组质粒后, 以泌乳期羊乳腺作为支持系统, 在处于稳定泌乳阶段分别注射不同量的重组质粒于山羊乳腺组织中。在注射后不同时间采集奶汁, 检测纤溶活性。
结果蚓激酶的不同重组质粒在注射到乳腺组织之后迅速得到表达, 注射后6~9 h蚓激酶表达量达到高峰, 并可持续至72 h以上; 不同表达载体间表达量略有变化, 但差异无统计学意义。
结论泌乳乳腺组织可以作为真核基因的1种快速有效的表达系统。

Abstract:
Objective To demonstrate the possibility of lactating mammary gland as a rapid and effective expression system for eukaryotic genes.
Methods Three eukaryotic vectors, pICLK, pLLKSN and pIbLK, were construeted, by using ctytomegalovirus (CMV)immediate early promoter, SV40 early promoter and beta-casein promoter as regulatory element, respectively, and utilizing the lumbrokinase(LK)cDNA as gene of interest.The recombinant expression plasmids were large-scale made and different amounts of each were injected into lactating goat mammary glands.Milk were collected at different periods and assayed for fibrinolytic activity.
Results The lubrokinase was immediately expressed from the three different recombinant plasmids.The expression reaches summit at 6~9 h after injection and persists for up to 72 h.Although variation in expression level exityed between different vectors, the difference was not significant.
Conclusion The lactating mammary gland is a rapid and effective expression system for eukaryotic genes.

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