织锦巴菲蛤(Paphia textile)过氧化氢酶基因cDNA全长克隆、序列同源分析与组织表达     

武祥伟  李佳凯  谭茎  刘贤德 《海洋学报(英文版)》2016,35(8):65-73

Catalase is an important antioxidant protein that can protect organisms against various forms of oxidative damage by eliminating hydrogen peroxide. In this study, the catalase c DNA of Paphia textile(Pt CAT) was cloned using RTPCR and rapid amplification of c DNA ends(RACE). Pt CAT is 1 921 bp long and consists of a 5′-UTR of 50 bp, a 3′-UTR of 349 bp, and an ORF of 1 542 bp that encodes 513 amino acids with a molecular weight of 58.4 k D and an estimated isoelectric point of 8.2. Sequence alignment indicated that Pt CAT contained a highly conserved catalytic signature motif(~(61)FNRERIPERVVHAKGAG~(77)), a proximal heme-ligand signature sequence(~(352)RLFSYSDP~(359)), and three catalytic amino acid residues(H~(72), N~(145), and Y~(356)). Pt CAT also contains two putative N-glycosylation sites(~(34)NKT~(36) and ~(437)NFT~(439)) and a peroxisome-targeting signal(~(511)AQL~(513)). Furthermore, Pt CAT shares 53%–88% identity and 29%–89% similarity with other catalase amino acid sequences. Pt CAT m RNA was present in all tested organs, including the heart, digestive gland, adductor muscle, gonad, gill, and mantle, but its expression was highest in the digestive gland. High-temperature-induced stress produced two expression patterns of Pt CAT m RNA: first, an initial up-regulation followed by a down-regulation in the heart, digestive gland, and gonad and, second, consistent down-regulation in all other organs. These results demonstrate that Pt CAT is a typical member of the catalase family and might be involved in the responses to harmful environmental factors.  相似文献   

泥蚶HDAC1基因cDNA全长、内含子克隆及时空表达特征分析   总被引：1，自引：0，他引：1  

任付真  姚韩韩  董迎辉  周小龙  林志华 《海洋学报》2016,38(8):73-82

HDAC1作为HDACs家族中重要成员,可使组蛋白去乙酰化进而调节基因表达,在细胞分化和胚胎早期发育中起重要作用。本文利用SMART RACE技术克隆得到泥蚶HDAC1(Tg-HDAC1)基因的cDNA全长序列,并对其内含子进行扩增及不同组织、不同发育时期的定量表达。结果发现:Tg-HDAC1基因的cDNA序列全长为2275 bp,开放阅读框(ORF)1587 bp,编码528个氨基酸;Tg-HDAC1蛋白序列与斑马鱼、鸡、小鼠等的相似性都在80%以上,表明该蛋白氨基酸序列在物种进化过程中具有保守性;在Tg-HDAC1基因中扩增出13个内含子,均存在于开放阅读框中,且都遵循GT-AG原则;荧光定量PCR(qRT-PCR)分析结果显示,Tg-HDAC1基因在成体血液、内脏团、外套膜、鳃、斧足和闭壳肌6个组织中均有表达,而在足中的表达量最高,且与其余5组织中的表达量有极显著差异(P<0.01),说明它对该组织生长具有重要作用。不同发育时期的表达差异结果表明,Tg-HDAC1基因在担轮幼虫期表达量最高,显著高于其他发育时期(P<0.05)。  相似文献   

无乳链球菌sodA基因的克隆、序列分析及原核表达载体构建     

《广东海洋大学学报》2016,(3)

无乳链球菌(Streptococcus agalactiae)是引起我国南方罗非鱼链球菌病的主要病原之一,SodA是无乳链球菌重要的毒力因子之一。根据链球菌sod A基因保守区设计引物,采用PCR扩增sod A基因部分序列,反向PCR和巢式PCR扩增其侧翼序列,成功获得无乳链球菌sod A基因。无乳链球菌sod A基因序列全长为699 bp,含开放阅读框609 bp,可编码202个氨基酸,演绎的SodA蛋白主要由α螺旋和不规则卷曲构成。BLAST分析表明,该蛋白氨基酸序列与犬链球菌(S.canis)及副乳房链球菌(S.parauberis)相应蛋白的氨基酸序列相似性分别高达80%和79%。将sodA基因克隆至原核表达载体pET28a上成功构建了重组质粒p ET28a-sod A,导入Escherichia coli BL21(DE3),诱导表达的重组蛋白质分子质量约为24 ku,主要以包涵体形式存在于表达菌中。  相似文献   

哈维氏弧菌qnr基因的克隆及原核表达条件优化     

《广东海洋大学学报》2016,(1)

根据Gen Bank中公布的哈维氏弧菌qnr基因序列设计引物扩增哈维氏弧菌qnr序列,将其插入p ET-32a质粒,构建原核表达载体p ET32-qnr,并对诱导温度、时间、IPTG浓度等条件进行优化。结果表明,哈维氏弧菌qnr全长651 bp,编码216个氨基酸;重组蛋白优化条件为于28℃、IPTG浓度为0.05 mmol/L条件下诱导6 h。  相似文献   

黄鳍鲷生长激素cDNA的分子克隆和序列分析   总被引：1，自引：0，他引：1  

张殿昌  江世贵等 《广东海洋大学学报》2002,22(4):62-65

  相似文献   

Characterization of defensin gene from abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> and its deduced protein     

洪旭光  孙修勤  郑明刚  曲凌云  昝金东  张进兴 《中国海洋湖沼学报》2008,26(4):375-379

Defensin is one of preserved ancient host defensive materials formed in biological evolution. As a regulator and effector molecule, it is very important in animals＇ acquired immune system. This paper reports the defensin gene from the mixed liver and kidney cDNA library of abalone Haliotis discus hannai Ino. Sequence analysis shows that the gene sequence of full-length cDNA encodes 42 mature peptides （including six Cys）, molecular weight of 4 323 Da, and pI of 8.02. Amino acid sequence homology analysis shows that the peptides are highly similar （70% in common） to other insects defensin. Because of a typical insect-defensin structural character of mature peptide in the secondary structure, the polypeptide named Haliotis discus defensin （hd-def）, a novel of antimicrobial peptides, belongs to insects defensin subfamily. The RT-PCR result of Haliotis discus defensin shows that the gene can be expressed only in the hepatopancreas by Gram-negative and positive bacteria stimulation, which is ascribed to inducible expression. Therefore, it is revealed that the Haliotis discus defensin gene expression was related to the antibacterial infection of Haliotis discus hannai Ino.  相似文献   

Efficient expression of green fluorescent protein （GFP） mediated by a chimeric promoter in Chlamydomonas reinhardtii     

吴锦霞  胡章立  王潮岗  黎双飞  雷安平 《中国海洋湖沼学报》2008,26(3):242-247

To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii, a high efficient expression vector was constructed. Green fluorescent protein （GFP） was expressed in C. reinhardtii under the control of promoters： RBCS2 and HSP70A-RBCS2. Efficiency of transformation and expression were compared between two transgenic algae： RBCS2 mediated strain Tran-Ⅰ and HSP70A-RBCS2 mediated strain Tran-Ⅱ. Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2, and the expression efficiency of GFP in Tran-Ⅱ was at least double of that in Tran-Ⅰ. In addition, a threefold increase of GFP in Tran-Ⅱ was induced by heat shock at 40℃. All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C. reinhardtii.  相似文献   

河口分汊的定量表达及其基本模式   总被引：3，自引：0，他引：3  

金元欢 《地理学报》1990,45(1):56-67

本文归纳出河口汊道间的四种交接方式:叉、合、结和口以及相应的十三种联线。并引入拓扑学中有关概念,定量表达点和线间关系。在分析河口汊道体系的拓扑学性质后,重新定义再结合因子,指出其与各种点和线、河口三角洲面积、河口汊道体系的外角等,均有一定的相关关系。在天然河口汊道体系中,最多出现的是点和口,其次是合和结;最多出现的线是fj,ff和fo,最少出现的是kk和ko线。最后,根据成因、形态和冲演特性相结合的原则,将河口分汉分成四个基本模式;少汊型、多汊型、河网型和游荡型分汊河口。并运用分汊河道体系的定量表示法,分析了各分汊模式河口汉道体系的拓扑学性质。  相似文献   

统计地震学进展：个人的体验     

D.  Vere-Jones  王晓权 《世界地震译丛》2007,(1):14-20

叙述了作者在发展统计地震学(作为一个描述、认识和预测地震的工具)方面的体验。集中讨论了地震发生随机模型的公式表达和使用。  相似文献   

角蛋白酶基因工程研究   总被引：2，自引：0，他引：2  

韩晓宇  刘亚洁  李江 《华东地质学院学报》2007,30(3):287-290

角蛋白是一种广泛存在于自然界中的硬性蛋白,主要存在于各种动物的毛发、鳞片、羽毛、蹄、角等结构中。角蛋白化学结构稳定,不易被一般的蛋白酶降解,但作为角蛋白酶的专一性底物,角蛋白可被角蛋白酶降解。角蛋白酶由微生物产生,可特异性降解羽毛等含角蛋白类的物质,在食品、医药、饲料生产和环境废物处理等方面具有潜在的应用价值。综述了微生物角蛋白酶基因分离和表达研究进展,并对角蛋白酶基因在异源表达过程中出现的问题进行了讨论。  相似文献