CHO表达生长激素释放因子的生物活性测定  

Determination of the Activity of Growth Hormone Releasing Factor Expressed in CHO Cells

作　　者：任晓慧[1] 刘松财[2] 张永亮[2] 欧阳红生[2] 赵建军[2] 张玉静[2] 

机构地区：[1]河北农业大学水产学院,河北秦皇岛066003 [2]解放军军需大学基础部,吉林长春130062

出　　处：《吉林农业大学学报》2003年第6期672-673,共2页Journal of Jilin Agricultural University

基　　金：国家自然科学基金(39500105)

摘　　要：在对生长激素释放因子(GIF)基因改造和化学合成,并构建了其真核表达载体pCDNA3-GRF(1-32)的基础上,用Lipofectin将上述载体转染CHO细胞进行瞬时表达,采用体外单层垂体细胞培养的方法对表达的CRF(1-32)进行了体外生物活性测定,即先将垂体用酶进行消化,再将消化细胞培养,形成单层细胞,利用其能合成与释放生长激素的能力来检测GRF的活性。结果表明:表达产物可刺激生长激素释放,并且比对照组提高3.8倍。Chemical synthesis of the GRF(1-32) gene had been finished in our former work. GRF(1032) gene, synthesized in this laboratory previously, was inserted into a plasmid , pcDNA3, to construct an expression vector in CHO cells. The expression vector was purified with a High Pure Plasmid Extraction Kit and then trasfected into prepared CHO cells in 60% confluent. Dispersed rat pituitary was used to determine the activity of expressed GRF(1-32) . The results showed that the expressed products stimulated Growth Hormone(GH) secretion in pituitary cells at high levels, being 3.8 times higher than the control group.

关 键 词：生长激素释放因子 垂体单层细胞培养 CHO细胞 瞬时表达 生物活性 体外测定 生长激素 

分 类 号：Q575.11[生物学—生物化学]