利用烟草表达一种新型鲑鱼降钙素的初步研究

A preliminary study on the expression of a new type salmon calcitonin gene in transgenic tobacco

利用烟草表达一种新型鲑鱼降钙素，以期为利用植物系统表达鲑鱼降钙素的研究提供理论参考。首先构建含外源基因“pil-msCT”的植物双元表达载体p35S-2300-twin T—DNAs：pil-msCT：NOS—ter，然后通过农杆菌介导法转化烟草，获得卡那霉素抗性烟草128株，其中PCR阳性植株14株（阳性率10．94％）；Southern blotting分析表明外源基因已整合到烟草基因组中，拷贝数为2～8个；半定量RT—PCR结果显示。转基因烟草株间“pil-msCT”转录水平存在较大差异。

Salmon calcitonin （sCT） has been proved to be an effective therapeutic peptide for Osteoporosis. In this study, tobacco was transformed with a modified gene msCT so as to produce a new type of sCT. Firstly, the plant binary expression vector p35S-2300-twinT-DNA ： pil-msCT ： NOS-ter containing foreign gene ＂pil-msCT＂ was constructed; Then, the fusion gene ＂pil-msCT＂ was introduced into tobacco plants by Agrobacterium-medlated transformation. 128 kanamycin resistant tobacco plants were regenerated, in which 14 were confirmed by PCR analysis （positive rate 10.94 % ）; Southern blot analysis proved that the copies of gene pil-msCT in tobacco plants ranged from 2 to 8 ; Semi-quantitative RT PCR detection showed different transcript levels of foreign gene among independent plants of To generation.