共查询到18条相似文献,搜索用时 140 毫秒
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为了研究串联抑制素基因免疫对绵羊孪生的影响,以猪抑制素α(1~32)基因及绵羊补体C3d基因作为选择摹因,利用RT-PCR技术构建串联抑制素重组质粒,并对60只绵羊进行免疫试验.结果表明:串联抑制素重组质粒pcDNA-DPPISS-DINH和pcDNA-DPPISS-DINH-sC3d3构建正确,并在BHK-21细胞中获得了分泌型表达.重组质粒pcDNA-DPPISS-DINH和pcDNA-DPPISS-DINH-sC3d3免疫绵羊后,双羔率分别为12.5%和25.0%,均与对照组间差异显著(P<0.05).串联抑制素重组质粒的成功构建,为抑制素基因疫苗的研制提供了理论依据和技术支撑. 相似文献
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为构建卵泡抑制素与绿色荧光蛋白(GFP)融合基因疫苗pEGISI,将pcISI中的乙肝表面抗原(HBsAg)S基因及插入S基因中的抑制素(INH)基因片段酶切回收;PCR扩增pcISI中INH基因片段,调整阅读框,一起融合到pEGFP-N1中EGFP基因的5'端,构建ISI-EGFP融合表达质粒pEGISI。酶切和测序鉴定表明,重组质粒pEGISI构建成功。将pEGISI转染293T细胞,16h后检测到绿色荧光,48h检测到强烈荧光,说明融合基因在293T细胞获得高表达;ELISA检测证实,表达产物ISI-EGFP融合蛋白具有INH的抗原抗体反应原性。质粒pEGISI的成功构建及表达为抑制素基因免疫的机理及安全性研究奠定了基础。 相似文献
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为构建卵泡抑制素与绿色荧光蛋白(GFP)融合基因疫苗pEGISI,将pcISI中的乙肝表面抗原(HBsAg)S基因及插入S基因中的抑制素(INH)基因片段酶切回收;PCR扩增pcISI中INH基因片段,调整阅读框,一起融合到pEGFP-N1中EGFP基因的5’端,构建ISI-EGFP融合表达质粒pEGISI。酶切和测序鉴定表明,重组质粒pEGISI构建成功。将pEGISI转染293T细胞,16h后检测到绿色荧光,48h检测到强烈荧光,说明融合基因在293T细胞获得高表达;ELISA检测证实,表达产物ISI—EGFP融合蛋白具有INH的抗原抗体反应原性。质粒pEGISI的成功构建及表达为抑制素基因免疫的机理及安全性研究奠定了基础。 相似文献
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根据GenBank中报道的猪抑制素基因序列设计两对引物,克隆含有抑制素抗原决定簇基因片段α(1~32)的p1INH和p2INH,在这两个片段中都设有限制性内切酶XbaⅠ的酶切位点,通过XbaⅠ把plINH与p2INH串联起来。串联抑制素基因片段与pcDNA3.1载体通过EcoRⅠ、HindⅢ酶切位点连接后经过转化DH5a感受态细菌构建重组载体。以重组载体作为模板,以含有EcoRⅠ、HindⅢ酶切位点的引物进行PCR,产物经验证为含串联的抑制素基因。串联的抑制素基因产生的融合蛋白具有抑制素的免疫原性,且不会与体内其他蛋白发生免疫交叉反应。串联后的基因片段在单位体积内的抗原位点数增多,免疫效果也会加强。可以反馈性地抑制FSH的分泌,提高母畜的排卵数,提高产仔数。 相似文献
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本试验在间接ELISA的基础上建立了一种检测抑制素(inhibit hormone, INH)表位多肽疫苗抗体的方法并对ELISA试剂盒进行优化,为今后测定主动免疫后细毛羊体内INH表位多肽疫苗抗体提供理论参考。在前人研究的基础上,采用间接ELISA法测定血清中INH表位多肽疫苗抗体的含量,通过对不同试验条件的控制摸索出最佳试验条件,即以脱脂奶粉为封闭液,INH及GnIH以表位多肽疫苗抗体稀释度为20 000倍,最佳反应时间60 min,最佳显色时间为15 min。本试验建立了一种检测细毛羊体内INH及GnIH抗体的方法,为今后对INH表位多肽疫苗抗体的研究提供了参考。 相似文献
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ZHANG Ling-ling YUE Yao-jing FENG Rui-lin LI Hong-feng GUO Ting-ting YUAN Chao NIU Chun-e LIU Jian-bin SUN Xiao-ping HAN Ji-long LIU Shan-bo YANG Bo-hui 《中国畜牧兽医》2016,43(8):2156-2163
In this study, an indirect ELISA method was established to detect inhibin hormone (INH) epitope peptide vaccine antibody, it would provide oretical reference for the determination of Fine-wool sheep after active immune body INH epitope peptide vaccine antibody. On the basis of the predecessors, using indirect ELISA method to determinate serum INH epitope peptide antibody levels of sheep, and through control different experimental conditions to look for the best experimental conditions. Through explorating the experimental conditions, finally, the testing experiment conditions were determined, which was blocked solution with skimmed milk powder, INH and GnIH synthetic peptides dilution degrees for 20 000 times, the optimum reaction time was 60 min, the best color action time was 15 min. In this experiment, a kind of method to detection antibody in the body after INH active immune sheep was built, it would provide a reference for future research. 相似文献
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将50只大鼠随机分为5组,分别注射0.2(T1)、0.6(T2)、1g/L(T3)pEGISI,100μg空载体pEGFP-N1(C1)和100μL生理盐水(C2),以探讨在没有使用免疫佐剂的情况下,抑制素与GFP融合基因疫苗pEGISI免疫对大鼠卵巢和生殖激素的影响。研究结果,加强免疫能提高抗体P/N值,T3组在加强免疫期与对照组差异显著(P〈0.05);加强免疫第2周时T3组抗体阳性鼠比例达40%,与T1和T2组差异均显著(P〈0.05)。T3组卵巢大小和卵泡数均显著高于对照组(P〈0.05),但抗体阳性鼠大卵泡数与阴性鼠差异不显著(P〉0.05)。各剂量组促卵泡素和雌二醇均高于对照组,且在加强免疫第2周时T3组与对照组差异显著(P〈0.05);各剂量组孕酮含量与对照组差异均不显著(P〉0.05)。结果表明,在没有使用免疫佐剂的前提下,抑制素pEGISI基因免疫可产生抗体,促进卵巢发育和FSH分泌。本试验条件下100μg是最佳免疫剂量。 相似文献
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回顾近30年来国内外畜禽生殖活动的内分泌免疫调控技术的发展,从早期的免疫类固醇激素和抑制素使动物提前进入初情期,促进更多的卵泡发育和提高排卵数,对公畜则提高其精子生成数量;到现在根据携带Inverdale基因母羊具有高繁殖力的原理发展的免疫BMP15或GDF9提高羊排卵数和产羔数的技术。对禽类免疫催乳素或免疫血管活性肠肽VIP抑制催乳素的分泌,可以解决易就巢禽类的就巢性,提高其产蛋性能。而免疫促性腺素释放激素则可以抑制性腺的发育进行“免疫去势”,提高畜禽的生长速度、瘦肉率、饲料报酬并改善肉质。今后内分泌免疫调控技术的研究应该开发具有更高免疫原性的新型基因工程疫苗和简单实用的免疫程序,以便更好地应用于生产实际。 相似文献
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Immunization of goats against inhibin increased follicular development and ovulation rate 总被引:4,自引:0,他引:4
Sasaki K Medan MS Watanabe G Sharawy S Taya K 《The Journal of reproduction and development》2006,52(4):543-550
In the present study, two experiments were conducted to induce superovulation in goats using passive and active immunization against inhibin. In the first experiment, two groups of goats were given an intravenous injection of either 10 ml normal goat serum (control; n=6) or inhibin antiserum developed against [Tyro30]-inhibin alpha (1-30) (passively immunized; n=6) 48 h before treatment with PGF2alpha. In the second experiment, two groups of goats were immunized with inhibin vaccine (actively immunized; n=5) or Freund's adjuvant (control; n=5) followed by three booster immunizations at 4 week intervals. Blood samples were collected for determination of FSH, LH, estradiol-17beta, and progesterone. Ultrasonography was used to determine ovarian activity at PGF2alpha injection and ovulation rate one week after estrus. In both experiments, there was a significant increase in plasma FSH concentration compared with the controls. However, the pattern of the FSH levels was different between the passively and actively immunized goats. The numbers of follicles in passively and actively immunized goats (22.4 +/- 2.3 and 18.6 +/- 2.1, respectively) were significantly greater than those in the controls (2.6 +/- 0.4 and 2.3 +/- 0.4, respectively). In addition, the ovulation rate was greater in the immunized animals compared with the controls. Therefore, either passive or active immunization against inhibin could be used to induce superovulation in goats. 相似文献