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1.
旨在初步了解丝氨酸蛋白酶抑制剂基因在扩展莫尼茨绦虫生长发育中的作用。采用分子克隆获得扩展莫尼茨绦虫丝氨酸蛋白酶抑制剂serpin基因部分片段,应用MEGA软件对其进行进化分析。同时以beta-Tubulin基因作为内参基因,采用SYBR Green real-time RT-PCR技术检测该基因在扩展莫尼茨绦虫4个不同发育阶段即头节、幼节、成节、孕节中的表达差异,最终克隆获得749bp的serpin基因片段。进化分析表明,扩展莫尼茨绦虫serpin基因与多方棘球蚴serpin基因有较近的亲缘关系。标准曲线分析显示,serpin和beta-Tubulin基因的Ct值与阳性质粒的浓度均呈良好的线性关系,相关系数均大于0.99,熔解曲线分析表明,产物为特异的单峰,具有较高的特异性。同时试验结果显示,serpin基因在虫体各发育阶段中的表达丰度存在差异,从高到低依次为孕节、头节、成节、幼节。由此初步推测,此基因可能在扩展莫尼茨绦虫虫卵发育为六钩蚴的过程中发挥一定作用。 相似文献
2.
为了克隆和表达编码扩展莫尼茨绦虫无精症缺失(DAZ)基因,试验根据GenBank中扩展莫尼茨绦虫DAZ基因cDNA序列(登录号为GH291478)设计1对特异性引物,以扩展莫尼茨绦虫组织总RNA为模板,RT-PCR扩增DAZ基因,并克隆到pMD19-T载体中进行序列测定,构建重组质粒pET28a-DAZ,转化BL21(DE3)大肠杆菌感受态细胞,以异丙基-β-D-硫代吡喃半乳糖苷(IPTG)进行诱导表达;利用非变性聚丙烯酰胺凝胶(SDS-PAGE)分析表达产物,通过螯合琼脂糖凝胶FF亲和层析柱对重组蛋白进行纯化。结果表明:重组DAZ基因在大肠杆菌中高效表达,表达的融合蛋白分子量约为14 ku。 相似文献
3.
运用组织学和组织化学染色法,分别对自然感染贝氏莫尼茨绦虫的成年绵羊（感染组）与正常成年绵羊（对照组）的小肠各肠段的上皮内淋巴细胞、浆细胞、杯状细胞和嗜酸粒细胞的分布和数量变化进行了比较研究。结果显示：感染组小肠各段上皮内淋巴细胞、浆细胞、杯状细胞和嗜酸粒细胞数量均显著高于对照组,其中感染组十二指肠、空肠和回肠的上皮内淋巴细胞数量分别比对照组增加了169.11%、230.38%和233.42%（P〈0.01）;嗜酸粒细胞数量分别比对照组增加了116.78%、123.87%和164.51%（P〈0.01）;浆细胞数量分别比对照组增加了127.34%、72.97%和328.26%（P〈0.01）;杯状细胞数量分别比对照组增加了33.40%、41.42%和133.17%。对照组和感染组上皮内淋巴细胞和杯状细胞从十二指肠到回肠均逐渐减少,相反,对照组和感染组嗜酸粒细胞数量从十二指肠到回肠均逐渐增多,对照组小肠固有层浆细胞数量从十二指肠到空肠增加,空肠到回肠减少,感染组小肠固有层浆细胞数量从十二指肠到回肠均逐渐增多;对照组十二指肠、空肠、回肠的上皮内淋巴细胞、上皮内杯状细胞、浆细胞和嗜酸粒细胞均差异极显著（P〈0.01）;感染组十二指肠、空肠、回肠的上皮内杯状细胞、浆细胞和嗜酸粒细胞均差异极显著（P〈0.01）,感染组上皮内淋巴细胞空肠与回肠差异极显著（P〈0.01）,但十二指肠与空肠差异不显著（P〉0.05）。研究结果表明,贝氏莫尼茨绦虫感染成年绵羊后,成年绵羊通过特异性黏膜免疫细胞上皮内淋巴细胞增生加强细胞免疫水平,浆细胞增生加强体液免疫水平,同时还通过非特异性黏膜免疫细胞,嗜酸粒细胞和杯状细胞的增生进一步加强黏膜免疫水平以抵抗贝氏莫尼茨绦虫对绵羊的感染。可见绵羊可以通过黏膜免疫相关细胞增生加强局部免疫力以监视虫体免疫逃逸来抵抗寄生虫的感染。 相似文献
4.
Prevalence and seasonal incidence of larval and adult cestode infections of sheep and goats in eastern Ethiopia 总被引：1，自引：1，他引：0
A study on the prevalence and seasonal incidence of cestode parasite infections of sheep and goats was carried out in eastern Ethiopia for 2 years (May 2003-April 2005). During this period, viscera including liver, lungs, heart, kidneys and the gastro-intestinal tract were collected from 655 sheep and 632 goats slaughtered at four abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga. At the abattoirs the abdominal, thoracic and pelvic cavities as well as the muscle surfaces of all animals were visually examined for the presence of larval (cystic) stages of cestode parasites. The viscera were transported within 24 h to the parasitology laboratory of Haramaya University and were examined for larval and adult cestodes following standard procedures. The most prevalent metacestodes (larval cestodes) were Cysticercus ovis (Taenia ovis), Cysticercus tenuicollis (T. hydatigena) and hydatid cysts (Echinococcus granulosus). In sheep, the overall prevalence was 26% for C. ovis, 79% for C. tenuicollis, and 68% for hydatid cysts. Similarly, for goats, the corresponding prevalence was 22%, 53% and 65%, respectively. The difference between sheep and goats in prevalence of C. tenuicollis was significant. The high prevalence of hydatid cysts in both sheep and goats indicates that cystic echinococcosis/hydatidosis is a public health problem in these regions which requires implementation of control measures, including public health education, strict meat inspection and control of stray dogs. The results of the survey also implies that infections of small ruminants with these metacestodes are responsible for condemnation of substantial quantities of affected organs and muscles and therefore of direct economic importance. Intestinal infections with adult tapeworms of Moniezia expansa, Avitellina centripunctata and Stilesia globipunctata, and bile duct infections with Stilesia hepatica were also common in both sheep and goats. In sheep, the overall prevalence of these tapeworms were 61%, 20%, 24% and 39%, respectively. Similarly, the overall prevalence of these parasites in goats was 53%, 21%, 27% and 36%, respectively. 相似文献
5.
[目的]分离和鉴定扩展莫尼茨绦虫(Monieziaexpansa)新基因,为进一步研究该基因的功能奠定基础.[方法]构建扩展莫尼茨绦虫成虫cDNA文库,随机挑取重组阳性克隆进行测序,对部分序列进行引物步移法测序,获取其全长cDNA序列;采用生物信息学等分析技术对该cDNA序列进行开放阅读框(ORF)的寻找、编码氨基酸的推导、核苷酸和氨基酸同源性比较及蛋白质二级结构的初步预测.[结果]获得了1个扩展莫尼茨绦虫新基因蛋白激酶C相互作用蛋白,全长1 527 bp,编码447个氨基酸,CDS预测存在明显的BAR,PDZ结构域.编码蛋白的理论分子质量为50.173 3 ku,等电点为5.22.[结论]获得了扩展莫尼茨绦虫蛋白激酶C相互作用蛋白的全长cDNA序列,为该基因功能的试验性鉴定工作奠定基础. 相似文献
6.
Abstract Greenhouse and field experiments were conducted to study the preference of two scarabaeid beetles, Anomala cupripes Hope and A. expansa Bates, for soybean plants at various growth stages and the potential use of a trap cultivar to control these pests. Insects preferred to feed on 6–9‐week‐old plants compared to younger ones. This may be due to more foliage on the older plants, which is a source of food and a shelter for the insect from direct sunlight. Planting a cultivar with more foliage as a trap cultivar between rows of the agronomic cultivar attracted the beetles and reduced the infestation of the agronomic cultivar. The optimum distance to plant such a trap cultivar between rows of agronomic cultivar needs to be determined for each location. 相似文献
7.
为表达扩展莫尼茨绦虫(Moniezia expansa)的烯醇化酶(Enolase),本研究采用RT-PCR方法以M.expansa 组织总RNA为模板扩增enolase基因,将其克隆到原核表达栽体pET-28a中,并转化BL21 (DE3)感受态细胞,以IPTG进行诱导表达.SDS-PAGE结果显示表达的重组Enolase约为47 ku.Western blot分析表明,天然蛋白能够被重组表达产物免疫小鼠的血清识别,出现两条免疫印记条带,即天然蛋白以两种形式(约47 ku和40 ku)存在,并具有抗原性,为该蛋白功能研究奠定了基础. 相似文献
8.
Takashi KOBORI Manabu MIYAGAWA Kunihiro NISHIOKA Satoshi T OHKI Takeshi OSAKI 《Journal of General Plant Pathology》2002,68(1):81-88
Local symptom expression and systemic movement of Cucumber mosaic virus (CMV) in Tetragonia expansa, Momordica charantia and Physalis floridana were mapped to the amino acid at position 129 of CMV coat protein (CP), using pseudorecombinants, chimeric RNAs, a site-directed
mutant of RNA 3 and four strains of CMV : pepo-, SO-, MY17- and Y-CMV. Local and systemic symptoms caused by three strains,
pepo-, SO- and MY17-CMV, and those by Y-CMV differed in the three host species. The three strains expressed local chlorotic
spots at 24°C and systemic chlorotic spots and ringspots at 36°C, whereas Y-CMV developed local necrotic spots at 24°C but
no systemic symptoms at 36°C in T. expansa. In M. charantia the three strains caused systemic chlorotic spots, whereas Y-CMV caused local necrotic spots. The three caused systemic mosaic
and Y-CMV systemic necrosis in P. floridana. With pseudorecombinants combined with pepo- and Y-CMV RNAs, CMV RNA 3 was responsible for symptom expression and systemic
infection. Inoculation with Y-CMV RNA 1, RNA 2 and chimeric RNA 3s exchanged CP gene fragments between pepo- and Y-CMV showed
that NruI-XhoI fragment of CP was essential for symptom expression. Comparative analysis of the NruI-XhoI fragments revealed that only the amino acid at position 129 was common among the three strains but different from that of
Y-CMV. Inoculation with a point mutant constructed by substituting one nucleotide resulting in an amino acid change from Ser
to Pro at position 129 in Y-CMV CP verified the previous experiments. These results indicate that the amino acid at position
129 of CMV CP is the determinant for local symptom expression and systemic movement in the three host species. CMV CP containing
Ser at position 129 may induce resistant responses in these plants.
Received 29 June 2001/ Accepted in revised form 28 August 2001 相似文献
9.
贝氏莫尼茨绦虫与扩展莫尼茨绦虫的形态学比较 总被引：2，自引：0，他引：2
[目的]将染色制片的贝氏莫尼茨绦虫与扩展莫尼茨绦虫进行形态学观察与比较,以探寻其种间分类鉴定的新依据.[方法]采用苏木素染色法,将采集的绵羊莫尼茨绦虫染色制片,以节间腺为分类依据,将贝氏莫尼茨绦虫与扩展莫尼茨绦虫分类后,分别各选取10条进行形态结构测量与比较,同时对两种绦虫的生殖器官和节间腺进行了对比观察与测量.[结果] 两种莫尼茨绦虫的长度和宽度,头节、幼节、成节、孕节的长度及宽度,吸盘直径,卵巢、卵黄腺、生殖腔形状及大小均有差异.[结论]两种莫尼茨绦虫部分形态结构、生殖器官形态结构可以作为种间形态学分类的参考依据. 相似文献
10.
分离和鉴定扩展莫尼茨绦虫(Moniezia expansa)新基因,为进一步研究该基因的功能奠定基础。构建扩展莫尼茨绦虫成虫cDNA文库,随机挑取重组阳性克隆进行测序,对部分序列进行引物步移法测序,获取其全长cDNA序列;采用生物信息学等分析技术对该cDNA序列进行开放阅读框(ORF)的寻找、编码氨基酸的推导、核苷酸和氨基酸同源性比较及蛋白质二级结构的初步预测。获得了1个扩展莫尼茨绦虫新基因——引发酶蛋白,全长1269 bp,编码422个氨基酸,属于AE_Prim_S家族。编码蛋白的理论分子质量为47.1598 ku,等电点为4.83。获得了扩展莫尼茨绦虫反应结合蛋白的全长cDNA序列,为该基因功能的试验性鉴定工作奠定基础。 相似文献