[人粒细胞集落刺激因子基因组基因的克隆及其在转基因小鼠乳腺中的表达]

[Cloning of human G-CSF genomic gene and its expression in transgenic mice mammary gland].

作者信息

Lu Y F, Tian C, Deng J X, Xiao C Z, Ma Q J

机构信息

Institute of Biotechnique Academy of Military Medicine, Beijing.

出版信息

Yi Chuan Xue Bao. 1999;26(4):281-7.

PMID:10593016

Abstract

Human genomic DNA was used as template of PCR. 1.5 kb G-CSF genomic DNA was obtained using PCR amplification method. Sequence analysis showed that genomic DNA sequence of human G-CSF was correct. The vector of mammary gland expression was constructed and contained whey acid protein (WAP) 5' control region directed human G-CSF genomic DNA. In order to produce transgenic mice, 1200 fertilized eggs were microninjected using WAP-G-CSF fragment. Two male transgenic mice were obtained and identified using PCR method and Southern analysis. Integration rate of human G-CSF gene was 2.37% in mice. Foreign gene could also be identified in F1 and F2 transgenic mice. Expression levels of human G-CSF in transgenic mouse milk were 120-250 ng/ml.

摘要

以人基因组DNA作为PCR模板。采用PCR扩增方法获得了1.5 kb的G-CSF基因组DNA。序列分析表明人G-CSF的基因组DNA序列正确。构建了乳腺表达载体，其包含指导人G-CSF基因组DNA的乳清酸蛋白（WAP）5'调控区。为了制备转基因小鼠，用WAP-G-CSF片段对1200枚受精卵进行显微注射。获得了两只雄性转基因小鼠，并采用PCR方法和Southern分析进行鉴定。人G-CSF基因在小鼠中的整合率为2.37%。在F1和F2转基因小鼠中也能鉴定到外源基因。转基因小鼠乳汁中人G-CSF的表达水平为120 - 250 ng/ml。