人乳铁蛋白基因转化银耳的研究  

Transformation of Human Lactoferrin Gene to Tremella fuciformis

作　　者：朱坚[1] 饶永斌[1] 谢宝贵[1] 刘新锐[1] 孙淑静[1] 郑域茹[1] 陈东兴[1] 

机构地区：[1]福建农林大学菌物研究中心,福建福州350002

出　　处：《热带作物学报》2010年第7期1137-1142,共6页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金(No.30471210);福建省卫生教育联合攻关计划项目(No.WKL2005-2-009)资助

摘　　要：利用电穿孔转化法,把携带人乳铁蛋白基因(hlf)的银耳表达载体pCB-hlf导入银耳芽孢菌株Tr21,用PCR鉴定该基因阳性转化子,检测转基因菌株的GUS活性,并用Southern杂交检测hlf的插入位点。结果表明:电穿孔转化法的转化率可达到9.25×106个/μg质粒DNA;抗性筛选的阳性转基因菌株中均能扩增出与预期大小相符的hlf、GUS和Tnos序列,且GUS基因在转基因菌株中均有表达。Southern杂交结果表明,不同转基因菌株hlf的插入位点不同,有些菌株插入了2个或更多的拷贝。RT-PCR结果表明,外源基因在银耳中能正常转录成RNA。Expression vector pCB-hlf which harbored human lactoferrin （hlf） gene was transformed to Tremella fuciformis by electroporation. Transformation rate was higher than 9.25×10^6/μg plasmid DNA. Some transformants were confirmed by PCR for the sequence of hlf and GUS and Trios. Southern blot analysis demonstrated that hlf gene was integrated in different sites, some transformants had been integrated two or more copies of hlf gene. The result of RT-PCR showed that hlf gene which harbored in transformant genome was transcribed to RNA.

关 键 词：人乳铁蛋白基因 表达载体 银耳 电穿孔 转化 

分 类 号：Q939.96[生物学—微生物学]