摘要
利用农杆菌转化法,将组成型表达β1,3葡聚糖酶及几丁质酶基因的双价植物表达载体pBLGC转化优质甘蓝型油菜品种H165,并得到了抗卡那霉素(Kanamycin,Kan)的再生植株。我们对所得到的抗Kan的再生苗进行了初步分子生物学检测,结果表明,在K15(Kan15mg/L)培养基上的绿苗中有30%为PCR阳性植株,而在K25(Kan10mg/L)培养基上的绿苗有53%的阳性率。对部分PCR阳性的转基因植株进行了点杂交分析,结果均表现出较强的杂交信号,这初步说明外源基因已整合到油菜基因组中。转基因植株的活体接种油菜菌核病菌(Sclerotiniasclerotiorium)试验表明,部分转基因植株比对照显示较强抗病性。
By Agrobacterium mediated method,the cotyledonary petiole of good quality rape variety H165 was transformed with plant expression vector pBLGC which constitutively express β 1,3 glucanase and chitinase genes.We obtained some Kanamycin(Kan) resistant regenerational green shoots,with these shoots,PCR identification was conducted.Results showed that 30% green shoots which grew in medium of Kan 15mg/L and 53% green shoots in Kan 25mg/L had positive reaction.We also made dot blot analysis with those green shoots,some of them gave positive signal,indicating that the foreign genes had integrated into rape genome.Fungal challenge of these transgenic plants showed that some plants were much more resistant to Sclerotinia sclerotiorium than non transgenic control plants.
出处
《生物工程学报》
CAS
CSCD
北大核心
2000年第2期142-146,共5页
Chinese Journal of Biotechnology
关键词
几丁质酶基因
转基因油菜
抗菌核病
农杆菌介导
glucanase gene
chitinase gene
transgenic plant of oilseed rape
resistance of Sclerotinia sclerotiorium
A.tumefacien
mediated transformation.
