摘要： 将位于pCHIT质粒上、已证明在植物中对真菌确有抗性、由35S启动子驱动的几丁质酶chi5B基因,插入到适于农杆菌介导转化的pCAMBLA1304质粒的HindⅢ酶切位点,构建出pAHCGG新质粒。以pAHCGG质粒为供体,通过根癌农杆菌介导转化烟草,XGluc、荧光检测表明,gus和gfp基因均能在植物体内表达;PCR检测证明chi5B基因在植物体内整合,并获得了转pAHCGG基因的烟草植株。
关键词:
质粒构建,
GFP荧光基因,
chi5B基因,
pAHCGG,
农杆菌,
转化,
烟草
Abstract: The fungal disease resistant gene,chitinase gene chi5B driven by CaMV 35S promoter,wasst ickily inserted into Hind III site of the vector pCAMBLA1304 that is suitable for Agr obacter ium-mediatedDNA t ransformation. The new constructed plasmid pAHCGG included gus and gfp (reported),npt II (bacterialant ibiotic selection),hph (plant ant ibiot ic selection),and chi5B (fungal disease resistant) gene constructs. Theplasmid could be used by the researches not only on improving techniques of plant genet ic transformation,but a-lso on chi5B gene expression in transgenic apples.
Key words:
Vector const ruct ion,
GFP gene,
Chitinase g ene,
Genet ic t ransformat ion,
Tobacoo
中图分类号:
XIE Xiao-liang, ZHAO He, WEN Chun-xiu. Construction of Resistant Gene Vector pAHCGG and It. s Application on Tobacco Genetic Transformation[J]. ACTA AGRICULTURAE BOREALI-SINICA, doi: 10.3321/j.issn:1000-7091.2003.z1.015.