三角褐指藻Δ5去饱和酶基因的原核表达     

刘冬梅  马晓磊  朱葆华  李斯  于文功  杨官品  潘克厚 《海洋湖沼通报》2012,(1):116-122

Δ5脂肪酸去饱和酶(Δ5fatty acid desaturase)是合成花生四烯酸(AA)和EPA的关键酶。为了构建三角褐指藻Δ5脂肪酸去饱和酶基因(简称D5)的原核表达重组质粒,并实现在E.coli BL21(DE3)中的表达。本研究根据GenBank中三角褐指藻Δ5脂肪酸去饱和酶基因序列,设计引物,扩增该基因并插入到pET28a中,测序鉴定后,转化E.coli BL21(DE3)感受态细胞,IPTG诱导表达,SDS-PAGE分析表达产物。结果表明pET28a-D5构建成功,IPTG能诱导D5特异性表达。  相似文献   

Ras gene in marine mussels: a molecular level response to petrochemical exposure     

Lima I  Peck MR  Rendón-Von Osten J  Soares AM  Guilhermino L  Rotchell JM 《Marine pollution bulletin》2008,56(4):633-640

Mussels are susceptible to numerous toxicants and are often employed as bioindicators. This study investigated the status of the ras proto-oncogene in Mytilus galloprovincialis following petrochemical exposure. A M. galloprovincialis homologue of the vertebrate ras gene was isolated, showing conserved sequence in regions of functional importance and a high incidence of polymorphic variation. Mutational damage was investigated in mussels chronically exposed to the water-accommodated fraction of #4 fuel-oil (WAF), and in mussels collected along the NW coast of Portugal in sites with different levels of petrochemical contamination. A ras gene point mutation was identified in the codon 35 of one individual exposed to 12.5% WAF. No mutations were detected in mussels from the WAF control or environmental samples. This represents the first report of a ras gene mutation, experimentally-induced by petrochemical exposure, in an invertebrate species.  相似文献   

我国主产地温石棉可吸入纤维致鼠肺损伤与抑癌基因p53、p16的表达     

曾娅莉  崔琰  邹文蓉  马骥  董发勤  张青碧  邓建军  杨洁  高霑 《岩石矿物学杂志》2019,38(6):843-851

选取我国四大矿区主产地的温石棉,制备成粒径小于10μm的可吸入纤维,采用非暴露式气管滴注法染毒Wistar雄性大鼠,分别于1、3及6个月时称重并处死,迅速分离肺组织,观察肺形态并称量,计算肺脏器系数,并采用HE染色法利用光学显微镜观察肺组织病理改变,RT-PCR法测定p53、p16基因表达,探讨我国温石棉可吸入纤维的生物安全性。结果表明,四大主矿区的温石棉纤维粒度基本小于10μm,符合可吸入纤维粉尘标准,研磨前后纤维粉尘的结构和活性基团无破坏;染毒组Wistar大鼠体重明显减轻,肺重量显著增加,肺脏器系数变大;染毒组Wistar大鼠肺组织发生严重病理改变,出现不同程度的充血、水肿、炎性细胞浸润和纤维增生;染毒组Wistar大鼠肺p53、p16基因表达均降低。综合以上结果,发现我国四大主矿区温石棉的可吸入纤维粉尘均对Wistar大鼠肺组织呈不同程度和方式损伤。随着染毒时间延长,Wistar大鼠肺组织的抑癌基因p53、p16表达显著降低,推测我国四大矿区温石棉的可吸入纤维粉尘具有致肺癌的风险。  相似文献   

A Preliminary Genetic Investigation of Rastrelliger Kanagurta Based on Random Amplified Polymorphic DNA and Mitochondrial ND2 Gene     

Siti Azizah Mohd Nor  Abu Talib A  Mohd Ghows M A  Samsudin B 《湿地科学》2008,6(4)

In a preliminary investigation, Random Amplified Polymorphie DNA (RAPD) analysis and partial mitochon-drial ND2 gene sequencing were conducted to study the genetic variation of the Indian mackerel, Rastrelliger kanagurta along a 450 km stretch of its distribution on the west coast of Peninsular Malaysia. A total of 53 individuals from 6 popu-lations were analyzed using 4 RAPD primers and a sub-sample of 15 individuals was chosen for sequencing of partial ND2 gene. Comparison between the 2 markers revealed genetic structuring in the RAPD results but genetic homogeneity for ND2 gene. Based on the former there may be at least 2 genetically differentiated groups of Rastrelliger kanagurta a-long this stretch.  相似文献   

Expression analysis of a Na+/H+ antiporter gene PeNHX1 from Populus euphratica     

YuXia Wu  JianQuan Liu 《寒旱区科学》2009,1(6):0541-0548

  相似文献   

Molecular Cloning and Expression ofPoIR2, a Novel Gene Involved in Immune Response in Japanese Flounder （Paralichthys olivaceus）     

Shuo Li  Chunmei Li  Xubo Wang  Yanan Wang  Zhipeng Liu  Teng Zhai  Quanqi Zhang 《中国海洋大学学报(英文版)》2010,9(1):59-64

A novel immune-related gene was expressed in Japanese flounder (Paralichthys olivaceus) injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5’UTR, a 336 bp open reading frame (ORF) encoding 111 amino acids and a 556bp 3’UTR. Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S (NMS), but further alignment indicated they did not have NMS C-terminal conservational signature domain. So it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa signal peptide and was a secretory pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after infected with V. anguillarum. These results indicated that the PoIR2 might play some important role in Japanese flounder immune response system. This gene was named PoIR2 (P.olivaceus immune-related gene 2, GenBank accession number: EU224372). The mature PoIR2 peptide was expressed in BL21(DE3) pLysS using pET-32a(+) vector and a great part of the recombinant mature peptide existed as soluble type.  相似文献   

太平洋真宽水蚤(Eurytemora pacifica)热休克蛋白70(HSP70)基因克隆及在金属胁迫下的表达分析     

李彬  景斐  武敏敏  张建设 《海洋与湖沼》2017,48(5):1060-1070

热休克蛋白70(HSP70)是生物体内一种重要的机体与细胞保护性蛋白。本文利用RT-PCR以及RACE技术首次克隆获得太平洋真宽水蚤(Eurytemora pacifica)HSP70简称Ep.HSP70 c DNA的全长序列,序列全长为2252bp(KY807149),开放阅读框(ORF)长1947bp,编码649个氨基酸,5′端99bp,3’端206bp;预测蛋白分子量为70.81k Da,等电点为5.16,为一种亲水性蛋白,不存在信号肽及跨膜区,含有丰富的α螺旋结构(37.60%),β折叠(18.80%)。同源氨基酸序列比对发现,与其他甲壳动物的同源基因保守性较高,尤其是HSP70家族典型的结构位点序列在甲壳类动物中具有高度保守性。系统进化分析表明,太平洋真宽水蚤和安氏伪镖水蚤(Pseudodiaptomus annandalei)进化关系最近;桡足类种内同源性要高于虾蟹类,与虾类同源性高于蟹类。荧光定量数据分析表明,不同浓度铜、镉、锌胁迫下太平洋真宽水蚤HSP70基因表达水平具有显著的时间效应与浓度效应的特征,三种金属对Ep.HSP70抑制效应呈现CuCdZn的趋势。Ep.HSP70基因的成功克隆及金属胁迫下的表达分析为深入研究HSP70蛋白生物学功能具有重要意义。  相似文献   

Lack of population genetic structure in the marine nematodes Ptycholaimellus pandispiculatus and Terschellingia longicaudata in beaches of the Persian Gulf,Iran          下载免费PDF全文

Narjes Sahraean  Jelle Van Campenhout  Annelien Rigaux  Hadi Mosallanejad  Frederik Leliaert  Tom Moens 《Marine Ecology》2017,38(3)

We investigated genetic diversity and population genetic structure of two common benthic nematode species, Ptycholaimellus pandispiculatus and Terschellingia longicaudata, from sandy beaches in the area of Bandar Abbas (Iran), Persian Gulf. Based upon partial mitochondrial cytochrome oxidase c subunit 1 (COI) gene data, 17 and two haplotypes were found for P. pandispiculatus and Te. longicaudata, respectively. Analysis of molecular variance did not reveal a significant population genetic structure for either species. The absence of genetic structuring indicates substantial dispersal and gene flow in our study area. To assess the species structure of Te. longicaudata at a larger geographic scale, we compared 18S rDNA and COI sequences from Iran and the Scheldt Estuary in The Netherlands to ascertain whether they truly belong to the same species. Our data confirmed previous studies that Te. longicaudata likely constitutes a complex of multiple cryptic species, with one of these species having a (near) cosmopolitan distribution.  相似文献   

瘤背石磺表皮生长因子基因的克隆、结构及进化分析   总被引：2，自引：0，他引：2  

杨铁柱  沈和定  史艳梅  刘欣  吴欣  李杰 《海洋科学》2017,41(3):8-16

首次在瘤背石磺(Onchidium struma)中克隆得到一种新的表皮生长因子(EGF)的cDNA序列全长。EGF基因cDNA的全长为1158bp,命名为Os-egf1,其中开放阅读框长度为846 bp,编码一条包含281个氨基酸残基的多肽链。根据氨基酸序列比对和结构域分析结果发现其含有2个保守的EGF结构域和1个EGF-like结构域,每个结构域中均包含至少6个半胱氨酸残基,且形成CX7 CX4-5 CX10-13CXCX8 C结构,其结构域由C1~C3、C2~C4和C5~C6之间形成的3个二硫键维持,符合表皮生长因子家族及其相关蛋白的特征结构域,但其余氨基酸序列与现有相关基因差异较大,推测可能是一种新的EGF-like基因。利用MEGA6.0软件构建Os-egf1与EGF家族相关蛋白的系统进化树,表明EGF家族蛋白具有一定的物种特异性。  相似文献   

脂肪营养对封闭循环水养殖道氏虹鳟生长、相关代谢酶及基因表达的影响     

张静  李勇  孙国祥  赵宁宁  马骏  王顺奎 《海洋科学》2017,41(10):24-32

为探讨饲料脂肪营养对封闭循环水养殖道氏虹鳟(Oncorhynchus mykiss)生长、脂肪代谢及相关酶基因表达的影响,作者采用单因素设计,即4个脂肪水平——12%、15%、18%、21%(以Ⅰ、Ⅱ、Ⅲ、Ⅳ组表示),共4个处理组,每处理3重复,每重复25尾鱼。试验鱼为初始体质量(333.25±20.71)g的道氏虹鳟幼鱼,试验期77 d。结果表明,与后3组相比,Ⅰ组试验鱼增质量率显著提高17.20%~33.17%,Ⅰ和Ⅱ组饲料系数显著低于Ⅲ和Ⅳ组6.20%~20.56%(P0.05);脂肪合成酶类FAS活力随饲料脂肪水平升高而降低,脂肪分解酶类LPL、HL及L-CPTⅠ活力变化特征与之相反(P0.05)。其中,Ⅰ组FAS活力显著高于Ⅲ、Ⅳ组7.5%、8.7%;L-CPTⅠ活力显著低于后3组13.08%~16.35%;Ⅳ组LPL活力显著或极显著高于前3组18.8%~44.7%(P0.05或P0.01);I组肝脏FAS的mRNA表达量比后3组显著高48.29%~55.15%(P0.05);LPL呈相反特征,Ⅳ组显著高于前3组36.72%~113.59%(P0.05或P0.01);Ⅰ组肌肉FAS mRNA表达量显著高于Ⅳ组66.82%(P0.05);LPL表达丰度随脂肪水平升高先下降后显著上升(P0.05)。该结果特征与肝脏中该两种酶活力变化具有同步性。研究发现,随饲料脂肪水平变化,肌肉FAS mRNA表达量变化比肝脏更明显且规律性更强,而肝脏LPL mRNA表达量变化较肌肉更明显。本试验初步确定,在封闭循环水养殖模式下,330 g左右道氏虹鳟饲料蛋白水平在45%时,脂肪水平以不超过12%为宜。  相似文献