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本试验测定了350头F2代杂种猪(长白猪×蓝塘猪)的6个肉质性状和7个胴体性状,用PCR-RFLP方法检测钙蛋白酶Ⅱ(CalpainⅡ)基因型,分析猪CalpainⅡ基因型间肉质和胴体性状差异,发现AA基因型与GG基因型猪肉质性状的4个值均差异显著,猪肌肉系水力、pH、大理石纹AA基因型显著高于GG、AG基因型;剪切力AA基因型显著低于AG(P<0.05)、GG(P<0.01)基因型;而胴体性状中三点背膘厚除活体C点背膘厚AA基因型高于AG基因型外,AA基因型均低于GG、AG基因型。结果表明,猪钙蛋白酶Ⅱ的AA基因型对于肉质和胴体性状而言是有益基因型,同时也说明CalpainⅡ基因参与了调控肌原纤维蛋白的降解,与促进动物骨骼肌生长、提高瘦肉率、改善肉嫩度等方面有一定相关。 相似文献
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WANG Leming WANG Yurui ZENG Zixuan RAO Guoshun WU Zhengjiao JIN Weikun WANG Dongying 《中国畜牧兽医》2022,49(2):677-686
【Objective】 This study was intend to obtain cathepsin L1(rFgCat L1) specific monoclonal antibody and construct the double antibody sandwich ELISA.【Method】 Five BALB/c mice were immunized with 1 mg/mL rFgCat L1 protein for four times.Mouse splenocytes were isolated and fused with SP2/0 cells to construct hybridoma cells.Strong positive hybridoma cell lines were screened, 1×106 cells were injected intraperitoneally per mouse to prepare monoclonal antibodies.Antibody titer and antigenic epitope were detected using ELISA method, antibody subtype and specificity were identified using Western blotting method.The double antibody sandwich ELISA was constructed by combining the anti-rFgCat L1 polyclonal antibody, and its sensitivity and specificity were tested.The positive and negative critical value was screened by 20 negative sera with positive control, and the constructed double antibody sandwich ELISA was verified by 47 goat positive sera and 47 dairy cow positive sera.【Result】 After immunization, the antibody titers in serum of 4 mice were all more than 104.After isolated mouse with the highest immune response spleen cells were fused with SP2/0 cells total of 8 of them were positive cell lines were obtained after selective culture.5D5 and 7G6 were identified as strong positive strains with stable antibody secretion.After multiple subcloning screens and subcultures, the antibodies secreted in the cell supernatant were stable, with titers of 29 and 210 respectively, with ascites titers of 107 and 108.Western blotting and antibody subtype identification kits identified that the two antibodies were IgG1 type and the light chain was kappa type, both of which could specifically bind FgESP.According to the same antigen site was recognized by the two kinds of antibodies, the antigen titer of the two monoclonal antibodies were comparied, 7G6 was used as the coating antibody, and anti-rFgCat L1 was used as the enzyme-labeled secondary antibody.The optimized condition of method was that 7G6 was coated at a concentration of 2 μg/mL, the dilution concentration of anti-rFgCat L1 polyclonal antibody was 25 μg/mL, the dilution of Don-HRP-conjugated was 1∶4 000, 5% skimmed milk powder was selected as the blocking solution and the color development time was 25 min.The method was proved that could recognize the lowest antigen concentration of 0.625 μg/mL, also could specifically recognize antigen of Fasciola fasciatus.The constructed sandwich ELISA method was used for antigen detection of 47 dairy cow positive serum and 47 goat positive serum infective samples kept in the laboratory and the positive antigen rate were 72.3% and 78.7%, respectively.【Conclusion】 Anti-rFgCat L1 monoclonal antibody was successfully prepared and the double-sheet sandwich ELISA method for fascioliasis was constructed, which provided a good theoretical basis and material basis for the development of low-cost and rapid diagnostic kits. 相似文献
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由EST获得全长cDNA对于结构基因组学和功能基因组学都是至关重要的,cDNA末端快速扩增技术RACE是该领域中的重要研究方法.利用BD SMART RACE技术扩增编码分泌天冬氨酸蛋白酶SA76基因的3'末端,将其与哈茨木霉cDNA文库中的SA76基因的EST序列进行序列拼接,获得2019bp的全长cDNA序列,其开放读码框长1593bp,5'非编码区266bp,3'非编码区201bp,编码530个氨基酸,有信号肽.哈茨木霉天冬氨酸蛋白酶基因与玉蜀黍赤霉、粗糙脉孢菌、球毛壳菌天冬氨酸蛋白酶基因的同源性分别为53%, 37%, 36%.利用BD SMART RACE技术首次从哈茨木霉中克隆天冬氨酸蛋白酶基因,为验证SA76基因的功能奠定基础,为进一步研究蛋白酶的作用机制及生物防治功能提供依据. 相似文献
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寄生虫半胱氨酸蛋白酶核酸疫苗的研究进展与应用前景 总被引:1,自引:1,他引:0
就寄生虫半胱氨酸蛋白酶的一些生理生化与免疫学性质进行了概括,阐述了该酶与宿主免疫应答之间的作用关系,证明了该酶具有作为研制高效核酸疫苗的条件,同时综述了寄生虫半胱氨酸蛋白酶核酸疫苗的发展以及应用前景。 相似文献
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