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1.
新城疫病毒(NDV)ND-xx08毒株经10 d龄SPF鸡胚增殖后,提取其基因组RNA并反转录成cDNA,用NDV F基因特异性引物,经PCR扩增后获得与F基因预期大小一致的DNA片段。将NDV F基因片段克隆到pMD18-T载体上,并进行EcoR I和Hind III双酶切鉴定和测序鉴定。结果显示,ND-xx08毒株F基因片段的长度为1 662 bp,共编码554个氨基酸,F蛋白的裂解位点为112R-R-Q-K-R-F117,是典型强毒株氨基酸序列结构。将NDV ND-xx08株F基因的47 bp到420 bp序列与新城疫病毒基因型I至基因型Ⅸ毒株的相同序列绘制病毒基因进化树,显示ND-xx08分离株属于基因Ⅶe型。将NDV ND-xx08株F全基因与国内外发表的23株NDV F基因核苷酸序列和氨基酸序列的同源性比较分析,结果表明,其核苷酸序列的同源性在82.7%~97.8%之间,氨基酸同源性在87.5%~97.7%之间。 相似文献
2.
根据GenBank发布的几丁质酶基因序列设计PCR引物,从叶片cDNA中克隆到了番茄几丁质酶基因,并命名为LeCHI,基因在NCBI中的登陆号为FJ849060。测序结果表明,LeCHI编码区全长为762 bp,编码253个氨基酸。RT-PCR检测表明,LeCHI基因在根、茎、叶和花蕾中均有表达。序列比对结果表明,LeCHI编码的氨基酸序列与同科的马铃薯、辣椒有较高的同源性。 相似文献
3.
珙桐rbcL基因的克隆与序列分析 总被引：1，自引：0，他引：1
根据已知植物rbcL基因设计引物,以珙桐叶片总DNA为模板,PCR扩增目的DNA片段,克隆入pDM19-T载体.阳性克隆鉴定后进行测序,序列分析结果表明:该基因片断长为1266 bp,包括1031 bp的编码区序列,编码343个氨基酸;其编码区氨基酸与烟草、菠菜、玉米、甘薯、拟南芥、水稻、葡萄、地钱和葡萄柚等9个物种的同源性94.13%以上,并构建了它们间的亲缘关系树.该基因片断的预测晶体结构与菠菜的最相近,推测Lys86、Lys60、Lys179等残基对酶的活性影响较大. 相似文献
4.
以NCBI中的EST数据库为主要数据来源,以一系列生物信息学软件为工具,进行了超氧化物歧化酶（SOD）基因的电子克隆及生物信息学分析。结果表明：通过电子克隆方法获得了SOD基因的编码区全长序列,该预测的SOD蛋白质氨基酸序列与数据库中同类基因的蛋白质氨基酸序列具有极高的相似性,并且含有完整的保守结构域;电子克隆到的SOD基因编码的不是分泌蛋白,也不是膜蛋白,而是胞质蛋白。通过某个基因的一个EST序列采用电子克隆的手段进行基因全长的电子拼接是可行的。 相似文献
5.
灰斑古毒蛾核型多角体病毒几丁质酶基因及其分子进化 总被引：5，自引：0，他引：5
通过对构建的灰斑古毒蛾Orgyia ericae单粒包埋型核型多角体病毒(OrerSNPV)基因组DNA的EcoRI酶切片段质粒文库的测序分析,在EcoRI—J片段(6．4kb)中鉴定出了编码几丁质酶(ChiA)基因开放阅读框(ORF),chiA基因编码区由1695bp组成,编码564个氨基酸,预计蛋白质分子量为62kD。这也是首次在基因序列水平上研究OrerSNPV。将OrerSNPV ChiA氨基酸序列与其它已知的18种杆状病毒ChiA氨基酸序列联配比较,结果表明OrerSNPV与其它杆状病毒ChiA氨基酸有60．3—69．5％同源性,其中与BmNPV、CfDEFNPV和LdMNPV的同源性最高。根据氨基酸序列绘制的分子进化树表明杆状病毒chiA基因至少可以分为SlMNPV、GV和其它NPV三个分支,OrerSNPV与LdMNPV在进化树上关系最为接近。 相似文献
6.
根据已克隆的野桑蚕酚氧化酶原基因的cDNA序列,利用生物信息学工具对该基因编码蛋白质的部分性质进行了分析.结果表明:野桑蚕酚氧化酶原基因PPO1和PPO2编码蛋白质的分子量和等电点分别为78.703 kDa,80.075 kDa和6.28,5.62;氨基酸序列的C-末端均没有疏水跨膜区域,其基因编码蛋白质的三级结构分别具有12个α螺旋、16个β折叠和14个α螺旋、18个β折叠.对18个物种的基因进化进一步分析表明:野桑蚕与家蚕的亲缘关系最近,与其他物种的亲缘关系相对较远. 相似文献
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8.
以新牧1号苜蓿为研究材料,采用RT-PCR方法,设计特异引物,从新牧1号苜蓿中克隆获得MvDREB基因cDNA,测序结果该片段全长为651 bp,Genbank登录号为GU073286.生物信息学分析结果表明,该基因序列与紫花苜蓿核苷酸序列相似度达99%,编码216个氨基酸,MvDREB蛋白氨基酸序列分析比对显示该蛋白属于AP2/ERF家族的亲水性核蛋白,蛋白质分子量为24873.1,理论等电点pI为5.90,不稳定参数为45.07. 相似文献
9.
以玉米Ｆ１减弱表达基因的ｃＤＮＡ片段为探针，从建立的玉米ｃＤＮＡ的文库中分离到相应的ｃＤＮＡ克隆．经测序和序列同源分析证明，该ｃＤＮＡ编码６－磷酸果糖－２－激酶／果糖－２，６－二磷酸酶，其氨基酸序列与人类、大鼠、牛和鸡等动物肝脏相同基因对应区段的相似性分为为６５．７％，６３．４％，６３．７％和６１．２％． 相似文献
10.
在真核生物的生命活动过程中,只有占基因组DNA总量的10％-15％具有为氨基酸编码的功能,而其余的DNA序列没有编码功能。这些DNA包括基因之间的间隔区和内含子,中度重复序列和没有表型效应的转座子,高度重复序列,特别是占总DNA 1％-3％的卫星DNA。而且有些DNA序列都是非转录的,它们既不为氨基酸编码,义不能转录成RNA。这样人们就无法用已知基因的功能来解释真核基因组如此之大的DNA含量,这称为C值矛盾。 相似文献
11.
孙建平 《通化师范学院学报》2006,27(4):81-82
1引言黄曲霉毒素简称AFT,是黄曲霉的代谢产物,为一种有毒物质,易污染食品,有较强的致癌作用,对人及动物的影响较大,因此检测食物中黄曲霉毒素毒性大小有很重要的意义.目前实验中常用的检测方法是高效液相色谱法(HPLC)及间接竞争酶联免疫吸附法(ELISA).本实验利用的是鸡胚培养法 相似文献
12.
目的：研究IL-4mRNA和IL-4蛋白在哮喘大鼠CD34^＋细胞中的转录表达及孟鲁司特（montelukast,MK）对其表达的影响。方法：将SD大鼠随机分为3组：哮喘组、MK组和正常对照组。用卵白蛋白制备大鼠哮喘模型。应用双抗体夹心酶联免疫吸附试验测定血浆中IL-4和γ-干扰素（IFN-γ）浓度;用MiniMACS磁珠分选系统分离骨髓CD34^＋细胞;采用SYBRGREEN I荧光实时定量PCR法测定CD34^＋细胞中IL-4mRNA的相对表达量。采用免疫组化技术测定CD34＋细胞中IL-4蛋白的表达量。结果：哮喘组骨髓CD34＋细胞中IL-4mR．NA和IL-4蛋白的表达量高于其它各组（p〈0．01）;哮喘组除了IFN-1水平低外,IL-4浓度和嗜酸性粒细胞（EOS）绝对值都是三组中最高的（P〈0．01）;除哮喘组外,其余组的各项指标相近（P〉0．05）。IL-4mRNA表达量与IL-4浓度、Eos绝对值呈正相关（P〈0．01）,与IFN-γ浓度呈负相关（P〈0．01）。结论：哮喘大鼠骨髓CD34^＋细胞中IL-4 mKNA的表达增强;孟鲁司特可以下调IL-4mRNA的表达。可能成为其抑制哮喘气道炎症形成的重要机制之一。 相似文献
13.
柠檬酸在肉鸡日粮中占0.5%,试验结果比对照组增重提高6.1%,公斤增重成本下降7.4%,成活率提高8.6%,鸡新城疫抗体效价提高1倍以上,饲料报酬与经济效益得到了明显的提高. 相似文献
14.
Objective: To investigate the effect of interleukin-18 (IL-18) on immune response induced by plasmid encoding hepatitis B virus middle protein antigen and to explore new strategies for prophylactic and therapeutic HBV DNA vaccines. Methods: BALB/c mice were immunized with pCMV-M alone or co-immunized with pcDNA3-18 and pCMV-M and then their sera were collected for analysing anti-HBsAg antibody by ELISA; splenocytes were isolated for detecting specific CTL response and cytokine assay in vitro. Results: The anti-HBs antibody level of mice co-immunized with pcDNA3-18 and pCMV-M was slightly higher than that of mice immunized with pCMV-M alone, but there was not significantly different (P>0.05). Compared with mice injected with pCMV-M, the specific CTL cytotoxity activity of mice immunized with pcDNA3-18 and pCMV-M was significantly enhanced (P<0.05) and the level of IFN-γ in supernatant of splenocytes cultured with HBsAg in vitro was significantly elevated (P<0.05) while the level of IL-4 had no significant difference (P>0.05). Conclusion: The plasmid encoding IL-18 together with HBV M gene DNA vaccines may enhance specific TH1 cells and CTL cellular immune response induced in mice, so that IL-18 is a promising immune adjuvant. 相似文献
15.
洪建捷 《温州大学学报(社会科学版)》2004,17(2):40-42,62
没食子酸丙醑在鸡肉宠物食品中具有较理想的抗脂肪氧化功能，大豆蛋白和含亚硝基亚铁血色原色素也具有一定的抗脂肪氧化能力． 相似文献
16.
为分析LPS刺激前后对RIPK2基因甲基化的影响,试验采用凝胶电泳、亚硫酸氢盐、双荧光素酶报告系统和qRT-PCR方法分析LPS刺激前后RIPK2基因的甲基化模式及甲基化相关试剂对鸡RIPK2启动子活性及表达水平影响。结果表明：LPS刺激后鸡RIPK2基因甲基化水平从53.8%下降至15.4%;5-氮杂胞苷显著性提高鸡RIPK2启动子活性,促进基因表达;而CpG甲基转移酶M. SssI显著性抑制鸡RIPK2启动子活性,抑制基因表达。研究结果对提高家禽免疫和控制过度性炎症反应具有十分重要的现实意义。 相似文献
17.
Type-dependent differential expression of neuropeptide Y in chicken hypothalamus （Gallus domesticus）
Chen GQ Hu XF Sugahara K Chen JS Song XM Zheng HC Jiang YQ Huang X Jiang JF Zhou WD 《Journal of Zhejiang University. Science. B》2007,8(11):839-844
Neuropeptide Y (NPY) is one of the most important orexigenic agents in central regulation of feeding behavior, body weight and energy homeostasis in domestic chickens. To examine differences in the hypothalamic NPY between layer-type and meat-type of chickens, which are two divergent kinds of the domestic chickens in feeding behavior and body weight, we detected mRNA levels of NPY in hypothalamic infundibular nucleus (IN), paraventricular nucleus (PVN) and lateral hypothalamic area (LHA) of these two types of chickens using one-step real time RT-PCR. The meat-type chicken had more food daily (about 1.7 folds) and greater body weights (about 1.5 folds) and brain weights than the layer-type chicken at the age of 14 d. In the meat-type of chicken, NPY mRNA levels of the IN and PVN were significantly greater than those of the LHA, and were not significantly different between the IN and PVN. However, in the layer-type of chicken, NPY mRNA levels were significantly greater in the IN than those in the LHA and PVN, and were not significantly different between the PVN and LHA. In all these hypothalamic regions, the layer-type of chicken had significantly higher NPY mRNA levels than the meat-type chicken did. These results suggest the expression of NPY in the hypothalamus has a type-dependent pattern in domestic chickens. 相似文献
18.
Rong-fa Guan Fei Lyu Xiao-qiang Chen Jie-qing Ma Han Jiang Chao-geng Xiao 《Journal of Zhejiang University. Science. B》2013,14(10):896-902
Meat quality traits of four genotypes of Chinese indigenous chicken [Ninghai chicken (NC), frizzle chicken (FC), Ninghai xiang chicken (XC), and Zhenning loquat chicken (LC)] and one genotype of commercial broiler [Arbor Acres plus broiler (AAB)] were analyzed. The indigenous chickens were raised before the commercial chickens in order to achieve the same final processed days. Indigenous chickens of NC, FC, XC, and LC showed significantly higher inosine-5′-monophosphate (IMP) content, shorter fiber diameter, and lower shear force than those of AAB (P<0.05). In the indigenous genotypes, NC and FC had significantly shorter fiber diameters and lower shear forces than XC and LC (P<0.05), and NC and XC had a higher IMP content than FC and LC (P<0.05). Moreover, the indigenous genotype of LC significantly displayed the highest protein content (P<0.05) in the five genotypes of birds, and no significant differences of protein content were found between the other genotypes of NC, FC, XC, and AAB (P>0.05). The indigenous chickens from FC displayed the highest total lipid content in the five bird genotypes (P<0.05). Significant differences of pH, color values of L* and a*, and drip loss for the five genotypes of birds were also observed. In conclusion, there were significant differences in the meat quality traits of the bird breeds selected in this study, and the indigenous chickens, especially the NC genotype, produced better quality meat as far as the IMP content, fiber diameters, and shear forces were concerned. 相似文献
19.
We investigated the possibility of producing chicken alpha interferon (ChIFN-α) in transgenic plants.The cDNA encoding ChIFN-a was introduced into lettuce (Lactuca sativa L.) plants by using an agro-infiltration transient expression system.The ChIFN-α gene was correctly transcribed and translated in the lettuce plants according to RT-PCR and ELISA assays.Re-combinant protein exhibited antiviral activity in vitro by inhibition of vesicular stomatitis virus (VSV) replication on chicken embryonic fibroblast (CEF).The results demonstrate that biologically active avian cytokine with potential pharmaceutical ap-plications could be expressed in transgenic lettuce plants and that it is possible to generate interferon protein in forage plants for preventing infectious diseases of poultry. 相似文献
20.
Transfection and expression of exogenous gene in laying hens oviduct in vitro and in vivo 总被引：2，自引：0，他引：2
Gao B Sun HC Song CY Wang ZY Chen Q Song HQ 《Journal of Zhejiang University. Science. B》2005,6(2):137-141
To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken 相似文献