摘要
用新疆甜瓜无菌苗子叶块，通过根癌农杆菌介导法，将黄瓜花叶病毒外壳蛋白cDNA基因导入“新红心脆”等6个品种（系），经卡那霉素（Kanamycin，Km）筛选．获得大量Km抗性植株，其中部分Km抗性植株经Luis法检测，表达了胭脂碱合成酶活性。进一步经SDS－PAGE，DOT-Elisa，PCR特异扩增，Western－Blot等方法检测，结呆证明了Km抗性植株基因组中整合了CMV—CP基因，长度为1kb，并能有效表达．表达产物分子量为24kD，确为CMV的外壳蛋白，其表达量占细胞总蛋白的5％。Km抗性植株经常规育种选育9代，获得了稳定过传的抗病、优质甜瓜品系K3－19。
he cotyloden strips of six Xinjiang melon varieties (lines)' Xin Hongxincui' etc. were transformed with Cucumber Mosaic Virus Coat Protein (CMV - CP) gene by A. tumefacien-mediated method, and large amo unt of anti-kanamycin (kan) plant were obtainted. After test by Luis method,some anti-kan plants showed the activity of nopalin synthase. Further tests, such as: PCR, dot blot,western blot, etc,indicated that the CMV-CP gene had integrated in the genomic DNA of the anti-kan plants, and the expression of this 1 kb gene in plant produced a protein of 24 kDa, an expected size for CMV-CP. The expression amount was about 5% of total plant protein. After traditional breeding of 9generations, the new melon line K3 - 19 of high quality and disease-resistance of stable heridity was obtained from these anti-kan plants.
出处
《西北农业学报》
CAS
CSCD
1999年第1期3-6,共4页
Acta Agriculturae Boreali-occidentalis Sinica
基金
新疆科委重点科研项目
关键词
新疆
甜瓜
根癌
农杆菌导法
抗病
优质新品系
Xinjiang melon
Cucumber Mosaic Virus coat protein gene
Transgenic plant
Agrobacterium tumefacien-mdiated transformation
New melon line of high quality and disease resistance
