摘要
目的:构建携带人类酸性成纤维细胞生长因子基因(haFGF)的植物表达载体,并将其转化进根瘤农杆菌中。方法:采取目的基因改造及载体构建的方法,以期构建高效表达载体。首先,采用植物偏好的密码子重新合成全长的haFGF,其次引入对目的基因表达有一定促进作用的表达调控元件:Ω序列、Kozak序列及KEDL内质网定位序列;将构建的携带haFGF的表达载体,利用冻融法将目的基因转化进根瘤农杆菌中;将构建的基因载体进行PCR鉴定、酶切分析及DNA测序鉴定。结果:DNA测序结果表明重新设计合成的基因与预期结果一致,PCR和酶切鉴定结果证实了haFGF植物表达载体构建和转化成功。结论:获得了携带haFGF的根瘤农杆菌菌株,为日后转基因植物工作奠定了一定的基础。
Objective： To construct an expression vector carrying haFGF gene, and to transformate Agrobacterium tumefacien. Methods： The method of modifying aim gene and constructing vector was adopted to develop a vector with high efficacy. At first, changing the aim gene with plant partial codes, and then, inserting the control dements, such as Ω sequence, Kozak sequence and KEDL, into the transgenic plants to improve the expression level. Result： DNA sequencing confirmed that the sequence of the synthetic gene was identical with that of the designed gene. PCR and restriction enzyme analysis showed that the construction of haFGF expression vector and its transformation to Agrobacterium tumefacie cells were successful. Conclusion： A strain of Agrobacterium tumefacien with haFGF gene has been obtained. This study provides a foundation for the transgenic study of plants.
出处
《贵阳医学院学报》
CAS
2006年第4期301-304,共4页
Journal of Guiyang Medical College
基金
国家高新技术研究发展(863计划)计划资助项目(编号2001AA215131
2002AA2Z3318)
国家自然基金重点项目(编号30230370)
