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1.
根据以发表的大鼠心肌线粒体肌酸激酶肌模型亚基基因(sMiMi-CK)的cDNA序列,设计并人工合成一对特异性引物,以大鼠心肌总RNA为模板,反转录聚合酶链反应(RT-PCR)扩增出一段编码大鼠心肌sMiMi-CK的DNA片段(1.33kb),将该片段克隆到载休pUC19上并进行初步鉴定,然后进行测序分析,结果与国外以报道的序列完全一致,证明已得到sMiMi-CK基因,为今后sMiMi-CK基因的表  相似文献   

2.
玉米苹果酸脱氢酶基因的分离与结构分析   总被引:9,自引:0,他引:9  
以一个玉米(ZeamaysL.)杂种一代超亲表达的cDNA片段为探针,从玉米幼苗期cDNA文库中筛选到一个全长1287bp的cDNA克隆。序列分析表明,该cDNA编码细胞质苹果酸脱氢酶,推导的氨基酸序列与龙须海棠(Mesembryanthemum crystallium L.)及拟南芥(Arabidopsis thaliana(L.)Heynh.)同一编码基因的氨基酸序列同源性分别为90%和84%。这是禾谷类作物中首次克隆的编码细胞质苹果酸脱氢酶的完整基因。  相似文献   

3.
三角酵母D—氨基酸氧化酶基因的克隆,测序及表达   总被引:1,自引:0,他引:1  
利用跨越内含子的PCR技术,从三角酵母(Trigonopsisvariabilis)变种FA110中扩增得到D氨基酸氧化酶基因(daao),并通过TA克隆的方法将其克隆至pGEMT载体。序列测定结果表明,所得daao基因的5′端内含子已被删除,基因总长度为1071bp,它与Trigonopsisvariabilis的D氨基酸氧化酶同源性达983%,与Fusariumsolani和Rhodotorulagracilis的同源性分别是389%和308%。为提高表达水平,又将此基因转移至高表达载体pET28b上,在大肠杆菌BL21(DE3)中进行诱导表达。经IPTG诱导,目的蛋白的产生量可占菌体总蛋白量的46%,分子量约为38kD。D氨基酸氧化酶的活力可达802u/L。  相似文献   

4.
根据已报道的番茄花叶病毒L株系(ToMVL)序列人工合成引物,经RTPCR扩增并克隆了我国番茄花叶病毒分离物(ToMVS1)的外壳蛋白CP基因及3′端非编码区。序列测定结果表明,所得cDNA共长682个核苷酸,其中CP基因含480个核苷酸,编码158个氨基酸,3′端非编码区含202个核苷酸,其核苷酸序列与ToMVL株系具有99.5%的同源率。将该基因片段克隆到pGEMEX1载体中,转入E.coli后诱导表达,经Westernblot检测证明,该基因已在大肠杆菌中正确表达。这是我国首次报道ToMVCP基因序列。  相似文献   

5.
二种淡水微囊藻rDNA16S-23S基因间隔区的序列测定与分析   总被引:10,自引:0,他引:10  
本研究采用PCR及序列测定的方法,对我国淡水铜绿微囊藻有毒株和另一低毒的种类惠氏微囊藻(M574)rDNA16S-23S基因间隔区进行了序列的测定和分析,结果表明:rDNA16S-23S基因间隔区可以作为一个精细且稳定的指标,用一微囊藻的分类和鉴定。并从分子水平提出了同囊藻与惠氏微囊藻在种系有较近缘的关系,本文首次对微落属MicrocystisrDNA基因间隔区全序列作了报道。为微囊藻属的鉴定及系  相似文献   

6.
鸡肌球蛋白轻链激酶表达载体的构建   总被引:3,自引:0,他引:3  
鸡肌球蛋白轻链激酶(MLCK)在调节平滑肌细胞收缩中具有重要作用。用PCR产生构建所需的限制性内切酶Sal I位点,将鸡MLCK cDNA插入质粒pBKrsv中,构建成pBKrsv-MLCK,并通过酶切图谱、SalI位点序列分析得到证实。重组质粒在大肠杆菌中获得表达。  相似文献   

7.
Moricandia是十字花科中唯一的具C3C4植物的属。为向十字花科芸苔属(Brasica)作物中转移C3C4等重要性状提供基本信息,从可交配性和RFLP指纹图谱两个角度研究了Moricandia与芸苔属作物间的亲缘关系。结果表明:Moricandia与芸苔属作物间的可交配性很低,仅从M.arvensis×甘蓝型油菜的105枚培养子房中获得4个杂种苗。用23个甘蓝型油菜核DNA探针和4个甜菜线粒体DNA探针检测了芸苔属作物和MoricandiaC3C4植物间DNA限制性内切酶位点多态性。Moricandia与芸苔属物种间的同源性很高,其中白菜型油菜与M.nitens之间在核基因组上的相似性甚至大于它与同属的甘蓝型油菜间的相似性。MoricandiaC3C4物种与芸苔属作物在核DNA和细胞质DNA间的高度同源性,揭示了通过有性杂交或原生质体融合及染色体间重组的途径向栽培种转移重要基因的可能性。  相似文献   

8.
鸡平滑肌肌球蛋白轻链激酶在NIH 3T3细胞中的表达   总被引:2,自引:0,他引:2  
肌球蛋白轻链激酶(MLCK)在调节平骨肌细胞收缩过程中具有十分重要的作用。本言语通过将MLCKcDNA插到质粒pBKrsv中构建pBKrsv-MLCK,并转染至NIH3T3细胞中,DNA-PCR、RT-PCR和Western blot分析表达转染细胞可表达MLCK。活生分析表明所表达的MLCK具有生物学活性。为进一步研究MLCK在信号传导,调节平骨肌收缩等作用奠定了基础。  相似文献   

9.
鸡肌球蛋白轻链激酶表达载体的构建   总被引:1,自引:0,他引:1  
鸡肌球蛋白轻链激酶(MLCK)在调节平滑肌细胞收缩中具有重要作用.用PCR产生构建所需的限制性内切酶SalⅠ位点,将鸡MLCKcDNA插入质粒pBKrsv中,构建成pBKrsv-MLCK,并通过酶切图谱、SalⅠ位点序列分析得到证实.重组质粒在大肠杆菌中获得表达.  相似文献   

10.
应用谷实夜蛾核型多角体病毒(HzSNPV)DNA聚合酶基因HindⅢ/PstⅠ3596bp片段作探针,经Sourthernblot杂交,克隆了中国棉铃虫核型多角体病毒(HaSNPV)完整的DNA聚合酶基因,大小约为3.4kb。限制性内切酶分析表明,HaSNPVDNA聚合酶基因限制性内切酶图谱与HzSNPV相似。用双脱氧链终止法测定该基因部分核苷酸序列(805bp),推导出编码区206a。序列同源性比较显示,HaSNPVDNA聚合酶与HzSNPV之间具有高度的同源性;与LdMNPV、AcMNPV、BmSNPV、CfMNPV和OpMNPV也具有一定的同源性  相似文献   

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正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases  相似文献   

16.
Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.  相似文献   

17.
Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.  相似文献   

18.
Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.  相似文献   

19.
肝癌中HBV和HCV基因和抗原的分布及意义   总被引:1,自引:0,他引:1  
采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细  相似文献   

20.
The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.  相似文献   

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