碱蓬亲环素基因SsCyp1cDNA的克隆、原核表达和耐盐性相关分析-生物化学与分子生物学专业论文.docxVIP

II II NaCl 浓度分别为 1%、6%、7%、8%的 40ml 液体 LB 培养基（含 0.01mM IPTG 和 50mg·L-1 ampicillin ） 中 培 养 ， 测 定 其 OD600 值 。 结 果 表 明 , 重 组 菌 (pET-dute-1/ SsCYP-1)比对照菌( pET-Duet-1)更具有明显的生长优势，推测出盐地碱蓬 SsCYP-1 基 因在抗盐胁迫方面具有重要作用。 关键词：盐地碱蓬，亲环素，肽脯氨酰顺反异构酶，耐盐性 PAGE PAGE IV Abstract Soil salinization seriously harms the agricultural development in the world and endangers the survival of human beings, which becomes the major environmental problem. In order to develop salty soil resources of and enhance the crop production with the application of biochemistry and molecular techniques, the salt-tolerant gene are isolated and cloned from salt-tolerant crops and transformed into other high-yield crops with low salt tolerance, making them grow healthily under the high-salt pressure. Cyclophilins are distributed extensively in the cytoplasm and miscellaneous organelles with the PPIase activity, which can catalyze cis-trans isomerization process and accelerate protein folding accurately. The results indicated that the enzyme activity was closely related to salt resistance and the over-expressed cyclophilin strikingly enhanced their tolerance capacity of the salt stress. Suaeda salsa is a typical indicator of the saliferous fields. In this paper, the gene encoding cyclophilin SsCYP-1 was cloned and over-expressed in E. coli (Escherichia coli) and then the salt tolerance activity in the LB culture medium was analyzed. The main results in the paper were listed as follows: The molecular characterization of the SsCYP-1 gene The full-length SsCYP-1 gene was cloned and assembled from Suaeda salsa with the degenerate primers and RACE methods. The full-length SsCyP gene sequence was 875bp in size with the open reading frame (ORF) of 531bp in size. The ORF encoded 176 amino acids with the predicted molecular mass of 18.8 KDa. The isoelectric point was 7.67 with the high abundance of Gly and Lys, which owned 21 basic amino acids (K, R), 20 acidic amino acids (D, E), 54 hydrophobic amino acids (A, I, L, F, W