首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 156 毫秒
1.
采用RT-PCR方法从小麦品种豫教2号的发育籽粒中克隆出淀粉合酶III基因(starch synthase III, SSIII)部分cDNA片段(509bp) (GenBank No. EF466009),同源性比较结果显示,它与GenBank 上已报道的SSIII基因有高度同源性。以pWM101质粒为基础,构建了由35S启动子调控的SSIII基因的反义表达载体pWM101SSIII;另外,还以pFGC5941质粒为基础,构建了SSIII基因的RNAi干扰载体pFGC5941SSIII,这些载体的构建为研究此基因的功能打下了很好的基础。  相似文献   

2.
根据GenBank收录的CRC基因cDNA序列设计引物,以短角果荠菜(Capsella bursa-pastoris)为材料,通过RT-PCR扩增出与拟南芥CRC基因同源的全长cDNA,进行测序、比对及同源性分析.结果显示,克隆的该基因cDNA序列与报道的拟南芥CRC序列一致性达到93%,可以推断其为荠菜CRC基因的cDNA(cbCRC).以Ti质粒pWM101为载体,构建了由CaMV35S启动子调控的cbCRC基因植物表达载体pWM101-cbCRC, 采用根癌农杆菌滴注柱头法转化拟南芥,获得了转cbCRC基因的拟南芥植株.转基因拟南芥心皮果荚形态大小发生了一定的变化,说明荠菜cbCRC基因在拟南芥中的表达对拟南芥心皮形态和大小都产生了一定影响,但其并没有使拟南芥表现出荠菜短角果的形态.  相似文献   

3.
拟南芥TT12和AHA10基因分别编码MATE类型和H+泵类型的跨膜转运蛋白,介导原花青素单体向液泡的转运,对种皮色素的积累起重要作用。甘蓝型油菜是重要的油料作物,但缺乏黄籽基因型,现有黄籽材料表型不稳定,黄籽性状的分子机理不清,缺乏相关的分子育种。克隆了芸薹属TT12和AHA10基因家族的RNA干扰载体片段BTT12I和BA-HA10I,以基于pFGC5941而改造的植物RNA干扰基础载体pFGC5941M为骨架,构建了相应的RNAi载体pFGC5941M-BTT12I(简称pBTT12I)和pFGC5941M-BAHA10I(简称pBAHA10I),通过分子鉴定后转化农杆菌,获得工程菌株,有利于通过基因沉默技术研究TT12和AHA10基因在芸薹属种皮色素跨膜转运中的作用和机理,探索黄籽性状的分子育种。  相似文献   

4.
甘蓝型油菜Fad2与Fae1基因双干扰RNAi载体的构建   总被引:1,自引:0,他引:1  
将来源于甘蓝型油菜XY15的Fad2与Fae1基因编码区,长度分别为349bp和426bp的两个保守序列片段连接成807bp的大片段。然后分别以正反向插入到pFGC5941干扰载体查耳酮合成酶内含子的两端,构建成RNAi载体。以期在菜籽中转录后能有效抑制Fad2与Fae1两个基因的表达。所构建的RNAi载体最终序列全长3kb,经限制性内切酶消化、PCR扩增验证和序列测定,证明目标序列与GenBank数据库中的碱基序列一致,并且为正反向插入到pFGC5941载体上。通过农杆菌介导的油菜转化,已获得油菜抗性再生植株144个。  相似文献   

5.
水稻OsNCED3基因的RNAi载体构建   总被引:1,自引:0,他引:1  
冯光秀  陈惠 《生物学杂志》2012,29(1):47-50,58
水稻OsNCED3基因是水稻抗逆过程中重要的基因之一.以水稻中花10号幼苗为材料,提取基因组DNA.设计引物扩增区段cDNA并引入相应的酶切位点,以基因组DNA作为模板,进行RNAi-OsNCED3顺式和反式目的片段的PCR扩增.将PCR产物连接到pMD19-T载体上,经酶切和PCR检测后进行测序.测序结果表明:RNAi-OsNCED3顺式和反式目的片段均已正确的连接到pMD19-T载体上.然后将RNAi-OsNCED3顺式和反式目的片段通过酶切和连接,连接到含有发夹结构的质粒pFGC5941上.PCR及双酶切结果显示,构建的pFGC5941-OsNCED3即RNAi-OsNCED3载体结构完整.  相似文献   

6.
苎麻CCoAOMT基因cDNA反义转化模式烟草''WS38''   总被引:1,自引:0,他引:1  
苎麻咖啡酰辅酶A氧甲基转移酶(CCoAOMT)是其木质素合成过程的一种关键酶,运用克隆的该酶基因cDNA及植物表达载体pBI121、pWM101,分别构建了35S启动子控制的苎麻CCoAOMT基因反义cDNA基因质粒(pBI121-antiBnCCoAOMT)和cDNA全长表达质粒(pWM101-BnCCoAOMT),并通过根癌农杆菌介导法将其转化至模式烟草WS38,获得了转基因烟草.对转基因植株进行分子分析和组织学初步研究表明,转反义RNA基因植株叶柄木质素含量较野生烟草或转正义基因烟草叶柄木质素含量降低.说明运用反义RNA技术对CCoAOMT基因的表达进行基因工程调控,一定程度上可以对木质素的合成产生干扰,为获得低木质素或木质素组分改良的苎麻基因工程奠定基础.  相似文献   

7.
拟南芥中的SIP1基因编码的蛋白与拟南芥盐胁迫应答中的关键蛋白SOS2存在互作关系,而NAC1为拟南芥中介导生长素信号促进其侧根发生的蛋白。本研究中我们将SIP1基因和NAC1正义基因以及SIP1基因和NAC1反义基因分别整合到一个经改造的具有2个35S启动子的可用于双基因表达的载体pFGC5941S中,构建了两个双基因表达载体pFGC5941S SIP1 NAC1 sense和pFGC5941S SIP1 NAC1 anti。并将这两个载体通过农杆菌介导的方法转化到野生型拟南芥中,共获得15株转基因植株。对这些转基因植株进行盐胁迫实验发现,在含75mmol·L-1 NaCl的MS培养基上,相比于野生型,pFGC5941S SIP1 NAC1 sense转基因植株主根增长,侧根数量明显增多,而pFGC5941S SIP1 NAC1 anti转基因植株长势与野生型苗相似。由此我们推测可能只有当SIP1和NAC1同时过表达时,才会促进盐胁迫下拟南芥侧根的发育。  相似文献   

8.
双价抗虫基因植物表达载体的构建   总被引:13,自引:0,他引:13  
将蝎毒基因BmKITS和几丁质酶基因chitinase2个抗虫基因采用不同的启动子ubi或35S,连到2个高效的植物表达载体pWM101和pBI101中,2个重组表达质粒分别经过限制性酶切分析和PCR鉴定,实验结果表明2个含有双价抗虫基因的植物重组表达质粒均已构建成功.  相似文献   

9.
拟南芥中的SIP1基因编码的蛋白与拟南芥盐胁迫应答中的关键蛋白SOS2存在互作关系,而NACl为拟南芥中介导生长素信号促进其侧根发生的蛋白。本研究中我们将SIPl基因和NAC1正义基因以及SIP1基因和NAC1反义基因分别整合到一个经改造的具有2个35S启动子的可用于双基因表达的载体pF—GC5941S中,构建了两个双基因表达载体pFGC5941S-SIP1-NAC1-sense和pFGC5941S-SIP1-NACl-anti。并将这两个载体通过农杆菌介导的方法转化到野生型拟南芥中,共获得15株转基因植株。对这些转基因植株进行盐胁迫实验发现,在含75mnlol·L-1 NaCl的MS培养基上,相比于野生型,pFGC5941S-SIP1-NAC1-sense转基因植株主根增长,侧根数量明显增多,而pFGC5941S-SIP1-NAC1-anti转基因植株长势与野生型苗相似。由此我们推测可能只有当SIP1和NAC1同时过表达时,才会促进盐胁迫下拟南芥侧根的发育。  相似文献   

10.
胡萝卜软腐欧文氏茵甜菜亚种(Erwinia carotovora subsp.betavasculorum)EcbCSL101菌株具有很强胞外酶分泌活性,接种非寄主植物烟草引起过敏反应.Southern blotting结果表明EcbCSL101菌株中含有hrpN基因.PCR扩增含EcbCSL101完整开放阅读框的DNA片段并克隆到表达载体pET28a( )中.核苷酸序列分析表明,EcbCSL101菌株的hrpN基因的ORF为1113 bp,编码36.65 kD HarpinEcbCSL101蛋白(GenBank,DQ355519),与其它几种软腐欧文氏菌Harpin蛋白有较高的同源性.将含有hrpNEcbCSL101基因的重组质粒转化到大肠杆菌JM109(DE3)中进行表达,纯化后的HarpinEcbCSL101能诱导烟草发生过敏反应.  相似文献   

11.
目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。  相似文献   

12.
13.
14.
国内外蝗害治理技术现状与展望   总被引:3,自引:0,他引:3  
张龙 《昆虫知识》2011,48(4):804-810
本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...  相似文献   

15.
The synthesis and turnover of cerebrosides and phospholipids was followed in microsomal and myelin fractions of developing and adult rat brains after an intracerebral injection of [U-14C]serine. The kinetics of incorporation of radioactivity into microsomal and myelin cerebrosides indicate the possibility of a precursor-product relationship between cerebrosides of these membranes. The specific radioactivity of myelin cerebrosides was corrected for the deposition of newly formed cerebrosides in myelin. Multiphasic curves were obtained for the decline in specific radioactivity of myelin and microsomal cerebrosides, suggesting different cerebroside pools in these membranes. The half-life of the fast turning-over pool of cerebrosides of myelin was 7 and 22 days for the developing and adult rat brain respectively. The half-life of the slowly turning-over pool of myelin cerebrosides was about 145 days for both groups of animals. The half-life of the rapidly turning-over microsomal cerebrosides was calculated to be 20 and 40 h for the developing and adult animals respectively. The half-life of the intermediate and slowly turning-over microsomal cerebrosides was 11 and 60 days respectively, for both groups of animals. The amount of incorporation of radioactivity into microsomal cerebrosides from L-serine was greatly decreased in the adult animals, and greater amounts of the precursor were directed towards the synthesis of phosphatidylserine. In the developing animals, considerable amounts of cerebrosides were synthesized from L-serine, besides phosphatidylserine. The time-course of incorporation indicated that a precursor-product relationship exists between microsomal and myelin phosphatidylserine. The half-life of microsomal phosphatidylserine was calculated to be about 8 h for the fast turning-over pool in both groups of animals.  相似文献   

16.
Today, serious health problems as overweight and obesity are not just constricted to the developed world, but also increase in the developing countries (Prentice 2006, Ramachandram et al. 2002). Focusing on this issue, BMI and percentage of body fat were compared in 2094 schoolchildren from two cross-sectional studies from India and Germany investigated in 2008 and 2009. The German children are in all age groups significantly taller, whereas the Indian children show higher values in BMI (e.g. 12 years: Indian: around 22 kg/m2; German: around 19 kg/m2) and in the percentage of body fat (e.g. 12 years: Indian: around 27%; German: around 18-20%) in most of the investigated age groups. The Indian children have significantly higher BMI between 10 and 13 (boys) respectively 14 years (girls). Indian children showed significant higher percentage of body fat between 10 and 15 years (boys) and between 8 and 16 years (girls). The difference in overweight between Indian and German children was strongest at 11 (boys) and 12 (girls) years: 70% of the Indian but 20% of the German children were classified as overweight. In countries such as India that undergo nutritional transition, a rapid increase in obesity and overweight is observed. In contrast to the industrialized countries, the risk of overweight in developing countries is associated with high socioeconomic status. Other reasons of the rapid increase of overweight in the developing countries caused by different environmental or genetic factors are discussed.  相似文献   

17.
以白术(Atractylodes macrooephala Koidz.)二倍体组培苗为材料,对其四倍体诱导方法进行研究,共获得45个白术同源四倍体株系,为优良株系的选育提供了材料。此外,还分析比较了其中8个白术四倍体株系与二倍体的过氧化物酶同工酶(POD)的酶谱差异,发现四倍体各株系过氧化物酶同工酶谱比二倍体的均多了Rf0.310的谱带,且总过氧化物酶比活力也发生了很大改变,对探讨白术四倍体优良株系的生理生化机理具有一定的参考价值。  相似文献   

18.
The molecular characterisation of species and genotypes of Cryptosporidium and Giardia is essential for accurately identifying organisms and assessing zoonotic transmission. Results of recent molecular epidemiological studies strongly suggest that zoonotic transmission plays an important role in cryptosporidiosis epidemiology. In such cases the most prevalent zoonotic species is Cryptosporidium parvum. Genotyping and subtyping data suggest that zoonotic transmission is not as prevalent in the epidemiology of giardiasis. Molecular characterisation of Cryptosporidium and Giardia is a relatively recent application that is evolving as new genes are found that increase the accuracy of identification while discovering a greater diversity of species and yet unnamed taxa within these two important genera. As molecular data accumulate, our understanding of the role of zoonotic transmission in epidemiology and clinical manifestations is becoming clearer.  相似文献   

19.
Enterococcus faecalis was the most frequently isolated enterococcal species from anal swabs and tonsils of dogs and cats, although in the anal samples from dogs Ent. hirae was found almost as often as Ent. faecalis. Most Ent.faecium strains from dog tonsils differed from those associated with humans and other animals in that they fermented sorbitol. Typical Ent. avium as well as atypical Ent. avium -like strains were seen in dogs, while the related species Ent. raffinosus was associated with cat tonsils. Enterococcus cecorum also occurred mainly in cats. Certain atypical strains, presumptively identified as Ent. cecorum , shared characteristics with Ent. columbae.
The most frequent streptococcal species in tonsils of cats and dogs were Streptococcus suis and Strep. canis. Streptococcus canis and Strep. bovis predominated in anal swabs. The canine Strep. suis differed from the common porcine strains in fermenting mannitol.
Forty-seven of the 288 isolates examined could not be identified or related to known species. The characteristics of two groups of these bacteria, provisionally called 'Ton 31 group' and 'O7 group' are described.  相似文献   

20.
The natural products novobiocin and derrubone have both demonstrated Hsp90 inhibition and structure–activity relationships have been established for each scaffold. Given these compounds share several key structural features, we hypothesized that incorporation of elements from each could provide insight to structural features important for Hsp90 inhibition. Thus, chimeric analogues of novobiocin and derrubone were constructed and evaluated. These studies confirmed that the functionality present at the 3-position of the isoflavone plays a critical role in determining Hsp90 inhibition and suggests that the bicyclic ring system present in both novobiocin and derrubone do not share similar modes of binding.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号