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1.
人乳铁蛋白基因在烟草中的表面及抗细菌与病毒的能力   总被引:6,自引:0,他引:6  
从人血液中性白细胞中分离细胞总RNA,用oligo(dT)为引物进行cDNA第1条链合成,再用乳铁蛋白cDNA的特异性此物进行聚合酶链反应(PCR),分两2段合成并拼成完整的乳铁蛋白cDNA,核苷酸序列测定证明,该cDNA所编码的氨基酸与前人论证的人乳铁蛋白的氨基酸序列完全一致,将该cDNA构建成植物表达载体p35ShLFc,经根癌农杆菌LBA4404感染烟草叶盘,获得具有卡那霉素抗性的烟草植株,  相似文献   

2.
生长素结合蛋白cDNA的克隆及其在烟草中的表达   总被引:1,自引:0,他引:1  
基于拟南芥内质网生长素结合蛋白基因的cDNA序列,设计合成了Ap5和Ap3两个引物,应用RT-PCR技术扩增了拟南芥的ABP基因。将该基因克隆在植物表达载体p35SSIN的35S启动子必Nos3‘端之间,得到植物表达载体p35SE。通过农杆菌介导的方法对烟草SR1进行了转化,由分子杂交等检测证明,生长素结合蛋白基因已在烟草中表达,同时转基因烟草后代对生长素的敏感性明显增加。  相似文献   

3.
甘蓝中BcpLH同源基因的克隆及其表达分析   总被引:3,自引:0,他引:3  
根据大白菜BcpLH基因设计引物,用PCR方法从甘蓝结球期的茎尖组织中克隆到了大白菜BcpLH基因的同源基因BoLH1。序列分析表明,BoLH1基因含有3个内含子,cDNA编码245个氨基酸,编码的蛋白中存在两个双链RNA结合蛋白结构域;Southern杂交结果显示在甘蓝基因组有1个以上的BoLH1基因拷贝。PCR表达分析表明,甘蓝BoLH1同源基因主要在结球期的茏尖组织、球叶和包叶中表达。推测B  相似文献   

4.
人乳铁蛋白基因在昆虫细胞中的表达   总被引:12,自引:0,他引:12  
将人乳铁蛋白cDNA(hLFc)克隆在质粒pBacPAK8的BamHI和SacI位点,构建成转移质粒8hLFc。该质粒DNA与AcNPV BacPAK6病毒株基因组DNA经共转染草地夜峨培养细胞Sf21,在培养的贴壁细胞中挑出空斑,经过3轮纯化,结合斑点杂交筛选,获得重组病毒。用蛋白质免疫印迹法检测到在重组病毒感染的Sf-21细胞中存在乳铁蛋白基因的表达产物。酶联免疫检测结果表明:重组人乳铁蛋白在  相似文献   

5.
采用cDNA-PCR技术,从人胎盘cDNA文库DNA中克隆了人碱性成纤维在子(hbFGF)基因,用PCR突变法对其5&端序列进行修饰,奖天然和修饰后的hbFGF基因分别克隆至表达载体pBV221,免疫抑迹和SDS-PAGE结果证明,经修饰后的基因在大肠杆菌DS5α中获得了表达,表达量占菌体总蛋白9%。  相似文献   

6.
根据已报道的马铃薯卷叶病毒基因组序列,设计合成一对特异性引物,以马铃薯卷叶病毒中国分离株的RNA为模板,反转录合成了cDNA第一条链,经PCR扩增后克隆于pUC19质粒中,进一步用PCR鉴定,限制酶切分析用序列分析。结果表明,PLRV-Ch56kD蛋白基因由1527个核苷酸组成,编码508个氨基酸,3‘端非编码区由141个核苷酸组成,与国外报道的苏格兰PLRV-S,荷兰PLRV-N,加拿大PLRV  相似文献   

7.
国内信息     
DEGCM技术中的RDA、SSH和RSDD差异表达基因分离技术 (DEGCM )发展很快 ,主要是PCR技术的发展和相应新技术的出现为DEGCM注入了新的活力。PCR引入差式筛选之中 ,可提高效力和降低假阳性率。若引入扣除杂交 ,可发展扣除扩增方案 ,并用特异引物结合于PCR中选择扩增差异cDNA而形成代表性差式分析法 (RDA)。采用抑制性PCR选择性扩增差异基因片段 ,用DNA链内“退火”优先于链间退火 ,使非目的序列产生“发夹”式互补结构 ,不能与引物配对 ,从而选择性抑制非目的基因片段的扩增 ,以此形成抑制性差…  相似文献   

8.
黑子南瓜甘油-3-磷酸酰基转移酶基因的克隆及序列分析   总被引:6,自引:3,他引:3  
依据国外报道的南瓜甘油-3-磷酸转酰酶(GPAT)基因的cDNA序列合成相应引物,用RT-PCR技术,成功地分离了黑子南瓜(Cucurbitaficifolia)GPAT基因的cDNA片段,并亚克隆到了pGEM-T载体系统的多克隆位点上,序列分析表明黑子南瓜GPAT基因的cDNA序列及递推的氨基酸序列与南瓜(Cucurbitamoschata)相比分别具有98%和965%的同源性。在1188bp中有22个核苷酸发生变化,导致13个氨基酸的改变  相似文献   

9.
以矮牵牛(Petunia hybrida L.)栽培品种为材料,取开放前的花蕾分离mRNA,反转录合成cDNA,以cDNA为模板,通过PCR扩增,对获得的目的片段进行序列分析。结果表明,分离的目的片段含有686个核苷酸(含有起始密码和终止密码)。核苷酸序列与文献报道相比,同源率为99.6%,只有3个碱基发生改变,5’端的MADS盒区域完全相同。将得到的矮牵牛花同源异型基因fbp2的cDNA(yfbp2)与CaMV355启动子和NOS3’终止子融合,构建了表达载体pBBP2。表达载体通过农杆菌(Agrobacterium tumefaciens)LBA4404(pAL4404)介导转化烟草(NicotianatabacumL.)叶片,在含有100mg/L卡那霉素的抗性培养基上再生成株。对抗性株进行总DNASouthern杂交和总RNA的点杂交,证明目的基因已导入烟草细胞中,整合到烟草基因组上,并且在烟草细胞中转录。同源异型基因fbp2导入烟草后导致烟草花型改变,在雄蕊上产生了花瓣。  相似文献   

10.
周雪平  刘勇 《病毒学报》1997,13(3):240-246
根据烟草花叶病毒U1株系序列,人工合成引物,用RT法合成了cDNA后,通过PCR技术扩增并克隆了烟草花叶病毒蚕豆株系的外壳蛋白的基因和3‘端非编码区。DNA序列测定结果表明,外壳蛋白基因全长480个碱基,编码158个氨基酸,3’端非编码区全长204个碱基,与TMV-U1株系的同源率为100%。  相似文献   

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It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.  相似文献   

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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   

17.
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   

18.
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   

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For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.  相似文献   

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