摘要
为研究在药用植物中表达人干扰素 (IFN) ,构建了人IFN -γ植物双元表达载体。方法是将IFN -γ基因的pstI片段插入克隆载体PBluescriptSK(+) 中 ,通过蓝白斑筛选以xbaI kpnI证实 ,以ndeI kpnI鉴定插入方向。得到中间载体PSKIFN,用xbaI kpnI双酶切PSKIFN ,回收 1.3kb片段 ,将此片段插入植物表达载体PROKII上 ,经限制性酶切分析及自动测序 ,结果证实插入片段大小及序列正确 ,构建获得成功 ,为进一步在药用植物中表达IFN
Plant binary expression vector of human Interferon-γ(HIFN-γ) was constructed to study the expression of in medicinal plant. The method was as follow. The plasmid of P SWIF including interferon-γ gene was digested with pst Ⅰ and low melting point agarose was used to recover the fragment of HIFN-γ.Then the recovered fragment was cloned to the vector of P Bluescript SK(+) to form an intermediate vector of P SKIFN .The P SKIFN was digested with kpn I /xba Ⅰ and then coned to plant expression vector of P ROK Ⅱ to form the plant binary expression vector of P ROKIFN . The P ROKIFN was identified with digestion of kpn Ⅰ / xba Ⅰ and automatic sequence system. The result of restriction analysis showed that the length of inserted fragment was about 1.3 kb and that of automatic sequence showed the sequence of inserted fragment was correct.
出处
《广州中医药大学学报》
CAS
1999年第3期194-197,共4页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
国家自然科学基金! ( 3 9770 90 2 )
国家中医药管理局! ( 97Z0 82 )
广东省科委科学基金 !( 960 5 4 2 )资助
关键词
干扰素Ⅲ型
转基因植物
药用植物
INTERFERON TYPE Ⅲ
PLANTS, TRANSGENIC
PLANTS, MEDICINAL
