山西瘦肉型猪(SD-Ⅲ系)基因组DNA的遗传分析(英文)

[目的]为评价山西瘦肉型猪(SD-Ⅲ系)的遗传特性提供相关参数,[方法]本文筛选了8条引物进行AFLP-PCR对30头猪进行了基因组DNA的分析。[结果]共获得206个AFLP标记,单引物获得的标记数在3～15个,群体相似系数为0.931(0.888～0.978)。[结论]AFLP适宜于基因组DNA遗传结构检测;SD-Ⅲ系猪纯度较高。     

牛Musclin基因片段克隆与序列分析

[目的]对牛Musclin基因片段进行克隆与序列分析。[方法]通过电子克隆技术克隆牛Musclin基因,并通过软件分析牛Musclin基因与小鼠、大鼠、人、猪和鸡的同源性。[结果]通过电子克隆技术成功克隆了牛Musclin基因;分析结果表明,牛Musclin基因与小鼠、大鼠、人、猪和鸡同源性分别为81．0％、80．8％、90．0％、89．1％和62．4％,预测的氨基酸序列含有“KKKR”结构和与小鼠ANP、BNP、CNP蛋白的同源性区域,并将克隆的牛Musclin基因片段注册GenBank（EF646361）。[结论]该试验为进一步研究Musclin在牛骨骼肌中的生物学功能提供了基础资料。     

猪肌肉生长抑制素基因的克隆及原核表达

本试验旨在通过基因工程方法获得重组肌肉生长抑制素蛋白。根据基因库肌肉生长抑制素基因序列设计合成特异性引物,引物两端分别加上EcoRⅠ和SalⅠ酶切位点及保护碱基,提取汉普夏猪的骨骼肌总RNA,用RT-PCR方法扩增肌肉生长抑制素全长基因,并将其克隆到pMD18-T载体上,测序后EcoRⅠ和SalⅠ双酶切,克隆到pET-30a( )中,构建原核表达载体pET-30a-M,将重组表达质粒转化至E.coliBL21(DE3)中,诱导表达。结果表明,成功扩增出肌肉生长抑制素全长基因;克隆载体经过DNA序列测定,所得基因与国外报道的完全一致;成功构建原核表达载体pET-30a-M;经IPTG诱导,表达出了6×His-M融合蛋白,用组氨酸标签抗体做Western印记证明产物大约48 ku,与预期大小相符,肌肉生长抑制素蛋白在大肠杆菌中成功的进行了表达。     

Cloning and Sequence Analysis of Lipoxygenase Gene cDNA from Cucumber Fruit （Cucumis sativus L.）

Lipoxygenases are nonheme-iron-containing dioxygenases that catalyze the hydroperoxidation of unsatrated fatty acids containing a cis, cis-1,4-pentadiene structure producing hydroperoxy acids with conjugated dienes. LOX activity has been found in a wide range of plants. Typical substrates for LOX in plants are linoleic acid and linilenic acid fatty acids. The function of various LOXs in plants is unknown, but their participation in all stages of plant growth and development has been suggested （Hildebrand, 1989; Siedow, 1991）. Some of the physiological processes in whicn lipoxygenses have been implicated include wounding （Saravitz and Siedow, 1996）, pathogen attack （Melan et al., 1993）, seed germination （Kato et al., 1992）, fruit ripening （Ferrie et al., 1994）, plant senescence （Paliyath and Droillard, 1992）. The study on the role of lipoxygenase in ripening and senescence fruit focused on tomato and strawberry. Cloning LOX gene of cucumber fruit will make us further understand the molecularaction mode of this enzyme during fxuit ripening and senescence. In this paper we isolated the partial nucleotide sequences of cucumber fiuit lipoxygenase gene and discuss the characterization of it.     

The first survey and molecular identification of Entamoeba spp. in farm animals on Qinghai-Tibetan Plateau of China

Protozoans of Entamoeba spp. are globally distributed zoonotic parasites that infect diverse animal hosts and humans. Prevalence and species/genotypes distribution of Entamoeba spp. in domestic animals are not fully investigated on Qinghai-Tibetan Plateau (QTP), an animal husbandry and agriculture region of China. In a survey, 528 fecal samples were collected from 7 species of domestic animals on multiple locations across QTP region and analyzed by PCR and sequencing analysis. The overall prevalence of Entamoeba spp. infection in all examined animals was 97.9 %. Four Entamoeba species, E. bovis, E. moshkovskii, E. ecuadoriensis and E. histolytica were found, and majority (94.2 %) of Entamoeba-infected animals harbored E. bovis. Six Entamoeba genotypes, Entamoeba ribosomal lineages (RL) 1, 2, 3, 4, 8 and 9 were identified by sequencing analysis. Two zoonotic species, E. moshkovskii and E. histolytica, were present in horses, while E. ecuadoriensis and E. bovis were found in horses and all species of seven farm animals, respectively. It was also observed that six Entamoeba genotypes were distributed in animals in specific pattern. The results revealed high prevalence of Entamoeba spp. infection in livestock, broad range of hosts as well as diversity and species/genotype distribution of Entamoeba spp. in farm animals inhabiting on the high altitude QTP region.     

JS/1/97/Go株GPMV NP基因的克隆、序列分析

根据GenBank中发表的GPMV SF02株的NP基因的核苷酸序列设计合成一对引物,采用RT-PCR扩增出与预期设计的1470bp大小相符的片断,将此扩增产物克隆入PMD18-T载体,进行序列测定和分析.结果表明:所扩增的NP基因的核苷酸长为1 470bp,共编码490个氨基酸.同源性分析表明:JS/1/97/Go与国内的SF02株的NP基因核苷酸同源性为98.6%,氨基酸同源性为99.6%.由此可见,鹅副粘病毒JS/1/97/Go分离株与SF02株的亲缘关系较近,同属于新城疫Ⅶ型基因.而其与LaSota株的核苷酸同源性为85.2%,氨基酸同源性为90.8%.说明该毒株相对于经典的NDV在NP基因上已发生了较大的变异.     

Genotyping of Mycobacterium bovis by geographic location within Mexico

The spacer oligonucleotide typing (spoligotyping) method was used to differentiate 62 Mycobacterium bovis isolates obtained from tissues with macroscopic lesions typical of tuberculosis in dairy cattle from different regions of Mexico. Our purpose was to see if a strain from one region was genetically different from those of other regions (with the long-term aim of doing molecular trace back of isolates obtained in the laboratory). Results from the genetic analysis indicate that M. bovis isolates cannot be grouped by geographic location due to a wide range of genetic types involved in dairy cattle infections. Isolates even from the same herd showed different spoligotypes but some isolates from different region had similar genetic patterns. Genetic typing without epidemiologic information does not seem to be a plausible method to trace back animals to source of origin to detect and eliminate sources of infection.     

番茄环斑病毒悬钩子分离物复制酶基因的克隆及序列分析

番茄环斑病毒(ToRSV)是一种高风险的检疫性有害生物,可侵染桃、李、葡萄、苹果、樱桃等多种植物。2000—2001年,我国从法国进口的葡萄插条上检出过此病毒。以ToRSV悬钩子分离物(Rasp-2)总RNA为模板,采用RT-PCR方法扩增病毒复制酶基因(RdRp)的cDNA片断并将其克隆到pMD18-T载体上。序列测定结果表明,Rasp-2RdRp基因由2133个核苷酸组成,编码711个氨基酸;Rasp-2半胱氨酸蛋白酶和RdRp之间的蛋白酶切割位点为Q/V。Rasp-2与其他分离物及株系RdRp基因的核苷酸序列同源性为79%～84%,氨基酸序列同源性为89%～94%。聚类分析结果显示,葡萄黄脉株系(GYV)与其他分离物及株系关系最远,Rasp-2与其他分离物及株系亲缘关系较近。证实Rasp-2与另外2个悬钩子分离物Rasp和Rasp-1的核苷酸序列存在明显变异。     

梨果实贮藏中可溶性果胶和半纤维素分子结构的变化

 为了了解分子结构变化与耐藏性的关系，以耐贮性不同的苹果梨和新高梨为试材，利用Ultrogel AcA 22凝胶柱渗漏分离，比较了梨贮藏中可溶性果胶和半纤维素分子大小的变化。结果表明，苹果梨的半纤维素分子大于新高梨。在贮藏过程中，耐贮性强的苹果梨果实可溶性果胶分子大小变化不明显，但半纤维素分子明显变小。相反，新高梨的可溶性果胶分子明显变小，而未观察到半纤维素分子大小的变化。说明梨的耐贮性与可溶性果胶分子结构变化及半纤维素分子大小有关，而与半纤维素分子结构变化没有直接关系。