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1.
增强玉米品种的抗虫性是减少玉米螟为害、提高玉米产量的有效途径。通过密码子优化改造获得抗虫基因Cry1A.301,构建原核表达载体pET30a-Cry1A.301,进行蛋白免疫学、ELISA检测以及玉米螟抗性生测。结果表明,原核表达载体中Cry1A.301蛋白高效表达,且7d后杀虫效率达100%。通过Cry1A.301基因与植物表达载体CPB连接,构建CPB-35S∶∶Cry1A.301-35S∶∶bar载体进行农杆菌介导的玉米萌动胚遗传转化,得到12株T0代转基因植株。T0代植株叶片中目的基因、筛选标记bar基因的PCR检测与蛋白免疫学检测结果表明,Cry1A.301基因已经整合到玉米基因组并表达。 相似文献
2.
李矮缩病毒外壳蛋白基因克隆及原核表达研究 总被引：1，自引：0，他引：1
为制备李矮缩病毒(Prunus dwarf virus, PDV)抗血清,克隆PDV外壳蛋白(CP)基因,构建原核表达载体,优化蛋白表达条件。以阳性感病甜樱桃叶片为试验材料,提取总RNA,根据PDV CP基因(L28145.1)设计特异引物,RT-PCR方法扩增,克隆、测序,构建原核表达载体pET30a-PDVCP,在大肠杆菌BL21(DE3)菌株表达。PDV CP基因（Genbank登录号：JF333587.1）全长657 bp,编码217个氨基酸,与GenBank其他PDV分离物CP基因核苷酸序列的同源性为89.2%~93.9%,推导的氨基酸序列同源性为97%~99%。根据完整CP基因核苷酸序列构建系统进化树显示：12个PDV分离物可分为3组,泰安分离物与巴西、土耳其等分离物属于Ⅰ组。成功构建原核表达载体pET30a-PDVCP,并在体外条件下诱导表达出融合蛋白。转pET30a-PDVCP载体的大肠杆菌BL21（DE3）菌株表达分子量约24 kDa的重组蛋白。该重组蛋白在30℃,1.0 mmol/L IPTG、诱导4 h表达量最大。 相似文献
3.
为制备樱桃病毒A(Cherry virus A,CVA)抗血清,克隆CVA外壳蛋白基因(CP),构建原核表达载体,优化蛋白表达条件。根据CVA全基因组序列(NC_003689.1)设计特异引物,RT-PCR方法扩增CVA外壳蛋白基因,克隆、测序,构建原核表达载体pET30a-CVACP,转化大肠杆菌BL21(DE3)株系,采用异丙基-β-D-硫代半乳糖苷(IPTG)诱导重组蛋白表达。序列分析显示,该区段长为603 bp,编码201个氨基酸,与法国分离物PF(HQ267856.1)的同源性为96.8%,与印度分离物JKSPMi-5(FN669548.1)同源性为86.3%。成功构建了原核表达载体pET30a-CVACP,SDS-PAGE电泳分析显示,转pET30a-CVACP载体的BL21(DE3)菌株表达分子量约24 kDa的重组蛋白,该重组蛋白在37℃、1.5 mmol/L IPTG、诱导4 h条件下表达量最大。 相似文献
4.
草莓轻型黄边病毒CP基因的克隆、序列分析及原核表达 总被引：2，自引：0，他引：2
克隆草莓轻型黄边病毒CP基因,并构建其原核表达载体,在大肠杆菌中诱导表达CP蛋白。利用SMYEV的检测引物,通过RT-PCR技术筛选带SMYEV的草莓植株;根据NCBI中SMYEV序列设计扩增CP基因全长引物,通过RT-PCR获得SMYEV沈阳分离物CP基因全长序列;将CP基因克隆到原核表达pGEX-6P-1上,利用IPTG诱导CP基因在大肠杆菌BL21(DE3)中表达蛋白。利用RT-PCR扩增获得SMYEV分离物SY05的CP基因全长序列,由729个核苷酸组成,编码242个氨基酸残基。SY05与其他SMYEV分离物的核苷酸同源性为80.1%～97.4%,氨基酸同源性为92.1%～99.6%,其中,与SMYEV分离物SY03的氨基酸一致性为98.3%。将SY05的CP基因成功克隆到原核表达载体pGEX-6P-1上,构建了重组表达载体p6P-EV-CP,并将其导入大肠杆菌BL21(DE3)中。SDS-PAGE分析表明,IPTG诱导SMYEV分离物SY05的CP基因在大肠杆菌中表达出分子量为52.0 kDa的融合蛋白。克隆出SMYEV分离物SY05的CP基因,实现了其在原核细胞中的表达,为SMYEV抗血清的制备奠定了重要基础。 相似文献
5.
摘要：克隆大肠杆菌乙酰辅酶A羧化酶（Acetyl CoA Carboxylase, ACCase）的生物素羧基载体蛋白（biotin carboxyl carrier protein,BCCP）亚基基因accB,构建accB的原核表达载体pGEX-4T-accB,转化大肠杆菌BL21（DE3）工程菌株, IPTG诱导进行融合表达,成功诱导表达了GST-accB融合蛋白,大小为43KDa。 相似文献
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K+通道蛋白在维持细胞离子平衡等生命活动中发挥重要的作用。利用真核或原核表达系统高效表达K+通道蛋白是深入研究其生化特征及生理功能的基础和前提。本研究在成功克隆巴西橡胶树(Hevea brasiliensis)K+通道蛋白基因cDNA全长的基础上,将HbKCO1 cDNA 编码区插入pET-28a构建原核表达载体pET-28a(+)-HbKCO1,并分别转化大肠杆菌RosettaTM(DE3) pLysS和BL21(DE3)菌株。经过条件优化,转化的大肠杆菌RosettaTM(DE3) pLysS菌株经1mM IPTG诱导3h能够高效表达HbKCO1重组蛋白,但同等条件下转化的BL21(DE3)菌株仅能微量表达。HbKCO1重组蛋白原核表达具有菌株依赖性。 相似文献
8.
内蒙古羊源细粒棘球蚴Eg95基因原核表达及蛋白鉴定 总被引：1，自引：0，他引：1
在克隆内蒙古羊源细粒棘球蚴疫苗候选基因Eg95的基础上,构建原核表达载体pET-Eg95并转化E.coli BL21(DE3),优化表达体系,获得Eg95纯化蛋白.将Eg95基因从克隆质粒pMD19-T-Eg95亚克隆到原核表达载体pET44a(+)上,构建原核表达载体pET-Eg95,转化E.coli BL21(DE3)诱导表达,优化表达条件.纯化的 Eg95 融合蛋白经 SDS-PAGE和Western Blot鉴定其正确性和免疫学活性.原核表达载体pET-Eg95在大肠杆菌中高效表达,优化的原核表达条件为:当菌液OD600值为0.6时,加入终浓度为0.1 mmol/L的IPTG,37℃,振荡培养5 h.纯化后的蛋白经SDS-PAGE和Western blot鉴定为目的蛋白并具有生物学活性.原核表达载体pET-Eg95构建正确并可高效表达可溶性Nus-Eg95融合蛋白,可作为特异性抗原应用于免疫印迹法检测血清抗体. 相似文献
9.
为进一步建立猪圆环病毒Ⅱ型(PCVⅡ)的有效检测方法,以及制备结构蛋白Cap和非结构蛋白Rep的单克隆抗体。根据GenBank中登录的PCVⅡ序列,设计2对特异性引物,PCR扩增出Cap、Rep蛋白基因,连接到pMD18-T simple质粒上,测序正确的Cap和Rep基因双酶切后分别插入到pACYCDuet-1上,构建成原核表达质粒pACYCDuet-1-Cap、pACYCDuet-1-Rep和pACYCDuet-1-Rep-Cap,然后将上述原核表达质粒转化BL21(DE3),并用IPTG进行诱导表达。结果表明：成功地在大肠杆菌BL21(DE3)中表达了猪圆环病毒Ⅱ型的Cap、Rep蛋白,且Rep、Cap蛋白共表达质粒pACYCDuet-1-Cap-Rep的表达效果优于其他2种质粒。 相似文献
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Jens Jensen 《Euphytica》1979,28(1):47-56
Summary The high-lysine gene in Risø mutant 1508 conditions an increased lysine content in the endosperm via a changed protein composition, a decreased seed size, and several other characters of the seed. The designation lys3a, lys3b, and lys3c, is proposed for the allelic high-lysine genes in three Risø mutants, nos 1508, 18, and 19. Linkage studies with translocations locate the lys3 locus in the centromere region of chromosome 7. A linkage study involving the loci lys3 and ddt (resistance to DDT) together with the marker loci fs (fragile stem), s (short rachilla hairs), and r (smooth awn) show that the order of the five loci on chromosome 7 from the long to the short chromosome arm is r, s, fs, lys3, ddt. The distance from locus r to locus ddt is about 100 centimorgans. 相似文献
12.
Hongyan WEI Jian ZHOU Jinguang LU Jingzheng SONG Qiongxi YU Zhihong JIANG 《Medicinal Plant》2019,(6):27-29
[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol... 相似文献
13.
Summary Apple and pear pollen was irradiated with doses of 0, 50, 100, 250 and 500 krad (gamma rays) and stored at 4°C and 0–10% r.h. From the in-vitro germination percentages an average LD 50 dose of about 220 krad was estimated. For both irradiated and untreated pollen a close and corresponding lineair relationship existed between germination percentage and pollen tube growth.Irradiated pollen was much more sensitive to dry storage conditions than untreated pollen, resulting in less germination and more bursting. Apparently, irradiation caused the pollen cell membrane to lose its flexibility faster than normal. Rehydration of dry-stored, irradiated pollen in water-saturated air restored germination percentages up to their initial levels. The importance of this procedure in germination trials is stressed. 相似文献
14.
Richard N. Lester 《Euphytica》1989,44(1-2):125-132
Summary An hypothesis is developed that the rapid change from wild plants into domesticated crops principally involves the selection of alleles with non-functional gene products which leads to reduced control of the highly integrated metabolism and morphogenesis previously accumulated by lengthy natural selection. Such disturbance of the genome produces altered physiological and morphological development which, although deleterious in nature, serves mankind better and has been selected. 相似文献
15.
B. Julier P. Guy C. Castillo-Acuna G. Caubel C. Ecalle M. Esquibet V. Furstoss C. Huyghe C. Lavaud A. Porcheron P. Pracros G. Raynal 《Euphytica》1996,91(2):241-250
Summary Twenty-five lucerne populations of the Medicago sativa complex, which were either diploid or tetraploid and wild or cultivated, were analysed for their resistance to four different fungal diseases and to stem nematode. Forage quality, including stem digestibility and saponin content, was also tested.Populations varied in susceptibility to the diseases caused by Colletotrichum trifolii, Verticillium albo-atrum, Sclerotinia trifoliorum and Pseudopezizza medicaginis, and to the nematode Ditylenchus dipsaci. Except for Sclerotinia rot, sativa and falcata subspecies differed in susceptibility, but this grouping of populations did not account for the full range of variation among them. However, the resistance to P. medicaginis was much lower in the sativa than in the falcata populations.Populations also varied significantly in stem fiber content and digestibility. Stem digestibility was negatively correlated to forage yield. Wild sativa and falcata populations had lower fiber content and higher digestibility than cultivated sativa populations. The medicagenic acid was the sapogenin responsible for the anti-nutritional effect of the lucerne measured by the yellow mealworm larvae Tenebrio molitor. The medicagenic acid content was lowest for the pure sativa populations, highest for the pure falcata populations, and intermediate for the French sativa varieties that have some traits originating from falcata germplasm. Some populations could be used in breeding programs to improve disease and nematode resistance, and forage quality. 相似文献
16.
In order to explore the pharmacological effects of active components of Gastrodiae Rhizoma on the central nervous system,through consulting related literatures,... 相似文献
17.
[Objectives]The purpose of this study was to investigate the effects of Pb on rhizosphere soil enzyme activity and chemical constituents of Achyranthes bidentat... 相似文献
18.
不同P-Zn配比对小麦幼苗微量元素营养的影响 总被引：2，自引：0，他引：2
采用了螯合-缓冲营养液培养方法对小麦进行了苗期培养试验,在3个P水平(0,0.6,3.0 mmol/L)和3个Zn水平(0,3,30 μmol/L)的完全组合下对小麦苗期生长及Zn、Fe、Cu、Mn营养进行了研究,旨在为小麦微肥施用提供理论依据.结果表明,P、Zn的正常供应促进了小麦生长,二者的缺乏与过量均会抑制小麦发育,且这种影响在冠部表现得更为明显.在小麦苗期,Zn与Cu的吸收存在明显的拮抗作用,但供Zn则促进了Zn和Cu的转运,而Mn转运则受到了抑制;过量供Zn时,大量Zn被转运到冠部,同时明显抑制了(Fe+Cu+Mn)的吸收总量;P的供应显著地抑制了Fe的吸收,但P的供应提高了Zn、Cu、Mn的转运率;P、Zn在对Zn与Fe、Cu、Mn间吸收竞争的影响中,Zn本身的影响要比P的影响更为明显,供Zn明显促进了小麦幼苗对Zn的吸收;在小麦幼苗冠部,Zn与Fe的竞争中,供P利于Zn的吸收,缺P则利于Fe的吸收;而Zn与Cu以及Zn与Mn间的竞争中,缺磷时利于Zn的吸收,供磷后则利于Cu和Mn的吸收.总之,小麦幼苗Zn、Fe、Cu、Mn营养中,P、Zn的不同配比会不同程度地改变Zn与Fe、Cu、Mn的协同或拮抗效应. 相似文献
19.
Summary
Glycine tabacina (Labill.) Benth. is a wild perennial species related to the cultivated soybean, G. max (L.) Merr. It is composed of diploid (2n=40) and tetraploid (2n=80) cytotypes. Currently, to differentiate the cytotypes, plants are grown out in the greenhouse and chromosome counts made on pollen mother cells. It is a laborious and time consuming process. The objective of this study was to determine whether electrophoretic techniques could be utilized to separate the cytotypes. Electrophoretic examination of seven isozyme systems from seed of 67 G. tabacina accessions revealed banding patterns that could be used to differentiate between diploid and tetraploid cytotypes in the species. Among the tetraploid accessions, the number of bands observed were always greater than the diploids. Some tetraploid banding patterns consisted of bands similar to the diploid tabacina and/or additional bands previously identified in other Glycine species. The patterns of isozyme multiplicity and variation in the tetraploid tabacinas suggests more than one mode of origin for the tetraploids. 相似文献
20.
Biological Activity and Quantification of Suspected Allelochemicals from Alfalfa Plant Parts 总被引：7，自引：0，他引：7
Autotoxicity restricts reseeding of alfalfa (Medicago sativa L.) after alfalfa until autotoxic chemical(s) breaks down or is dispersed into external environments. A series of aqueous extracts from leaves, stems, roots and seeds of alfalfa ‘Vernal’ were bioassayed against alfalfa seedlings of the same cultivar to determine their autotoxicity. The highest inhibition was found in the extracts from the leaves. Extracts at 40 g dry tissue l?1 from alfalfa leaves were 15.4, 17.5 and 28.7 times more toxic to alfalfa root growth than were those from roots, stems and seeds, respectively. A high‐performance liquid chromatography (HPLC) analysis with nine standard compounds showed that the concentrations and compositions of allelopathic compounds depended on the plant parts. In leaf extracts that showed the most inhibitory effect on root growth, the highest amounts of allelochemicals were detected. Among nine phenolic compounds assayed for their phytotoxicity on root growth of alfalfa, coumarin, trans‐cinnamic acid and o‐coumaric acid at 10?3 m were most inhibitory. The type and amount of causative allelochemicals found in alfalfa plant parts were highly correlated with the results of the bioassay, indicating that the autotoxic effects of alfalfa plant parts significantly differed. 相似文献