NPTⅡ基因在满江红鱼腥藻染色体DNAglnA位点的定点整合及对泌氨和细胞形态结构的影响  

Site Directed Integration of NPT Ⅱ Gene Into GlnA of the Chromosomes in Anabaena Azollae and its Effects on Morphology, Ultrastructure and Ammonia Secretion of the Transformed Cells

作　　者：吴晓强[1] 施定基[2] 刘祥林[1] 汪洪杰 邱泽生 张承谦 印莉萍[1] 赵微平[1] 

机构地区：[1]首都师范大学生物系 [2]中国科学院植物研究所

出　　处：《首都师范大学学报（自然科学版）》1993年第4期27-33,共7页Journal of Capital Normal University：Natural Science Edition

基　　金：国家自然科学基金;北京自然科学基金

摘　　要：本研究构建了含glnA基因片段和TPTⅡ基因的单交换整合型重组质粒pGN1-4,并通过电激法,三亲接合转移和二亲接合转移法号入到满江红鱼腥藻细胞中。用蓝细菌原位杂交法筛选出转化细胞菌落。再经Southern杂交方法进一步证实了转化结果。化学测氨法表明,培养后7～14d泌氨达到高峰,转化细胞泌氨比对照高40％。光镜观察表明,转化细胞藻丝变短,易断成单个细胞,异形胞频率降低;扫描电镜观察指出,转化细胞外壁有鳞片状鞘层,而对照较光滑。Suicide vector pGN1-4 were constructed from glnA in pAn503 and NPTⅡ in pRL447,which were succesfully introduced into Anabaena azollae cells by methods of electropora-tion and conjugation. The positive transformed colonies were screened out by in situ hy-bridization with probe of pRL447 and the results were futher proved by Southern hy-bridization. By optical microscope, scanning and transmission electrical microscope it wasobserved that cell structure and morphology of transformed Anabaena azollae strainsshown and obovious variance such as the filaments became shortened and were easily bro-ken into single cells by one cycle of freezing and thawing treatment. Compared with thesmooth shape of control cells it was also observed that there some redudant excretive sub-stance attached on the envelope of transformed cells. By chemical mehtod of ammoniameasurement, it was found that the amount of ammonia secretion from transformed cellswas 40% higher than those from control cells.

关 键 词：满江红鱼腥藻 NPTII基因 glnA 定点整合 泌氨 细胞形态结构 

分 类 号：Q342.[生物学—遗传学]