CAJ | 학술논문

对三孢布拉氏霉菌番茄红素环化酶(CarRA)进行抗原表位分析并制备兔多克隆抗体。运用GOLDENKEY分子生物学软件对三孢布拉氏霉菌CarRA结构蛋白的抗原表位及其二级结构进行了分析比较，结合现有的抗原表位选取原则从中筛选了一段显示表位特征的氨基酸残基序列，应用固相合成法合成三孢布拉氏霉菌 CarRA 多肽，然后将其与KLH 偶联制成免疫原，应用于动物免疫及多克隆抗体的制备，对该多抗与三孢布拉氏霉菌 CarRA 的反应性进行了检测。获得了三孢布拉氏霉菌CarRA的多抗，该抗体经间接酶联免疫吸附测定法检测其效价为1：32000，Dot blot检测在三个稀释度下都出现了目的条带，证实该抗体具有较好的反应性和特异性。成功制备了三孢布拉氏霉菌CarRA多抗，为进一步明确发酵促进剂的作用机制及其对三孢布拉氏霉菌代谢网络的影响，对CarRA进行定量以及对其相关功能进行深入研究提供了有利条件。
대삼포포랍씨매균번가홍소배화매(CarRA)진행항원표위분석병제비토다극륭항체。운용GOLDENKEY분자생물학연건대삼포포랍씨매균CarRA결구단백적항원표위급기이급결구진행료분석비교，결합현유적항원표위선취원칙종중사선료일단현시표위특정적안기산잔기서렬，응용고상합성법합성삼포포랍씨매균 CarRA 다태，연후장기여KLH 우련제성면역원，응용우동물면역급다극륭항체적제비，대해다항여삼포포랍씨매균 CarRA 적반응성진행료검측。획득료삼포포랍씨매균CarRA적다항，해항체경간접매련면역흡부측정법검측기효개위1：32000，Dot blot검측재삼개희석도하도출현료목적조대，증실해항체구유교호적반응성화특이성。성공제비료삼포포랍씨매균CarRA다항，위진일보명학발효촉진제적작용궤제급기대삼포포랍씨매균대사망락적영향，대CarRA진행정량이급대기상관공능진행심입연구제공료유리조건。
To analyze the epitope of Blakeslea trispora CarRA protein and prepare its polyclonal antibody. Combined with existing antigen epitope selection principle, the Blakeslea trispora CarRA full-length sequence was analyzed by using the bio-informatics software GOLDENKEY to detect the structure of amino acids and select its epitopes. The Blakeslea trispora CarRA peptides fragment was synthesized by solid-phase peptide synthesis method and purified by HPLC. The synthetic antigen was conjugated with the keyhole limpet hemoyanin (KLH), and then rabbits were immunized with this antigen and the antibody was examined. The results show that the antibody titer is as high as 1:32000. Through Dot blot analysis, it was proved that this peptide antibodies could react with Blakeslea trispora CarRA. All these prove that the antibody has wonderful reaction and speciality. A CarRA-specific antibody was succeddfully developed, which lays foundation for further studies of fermentation accelerator mechanisms, effects on Blakeslea trispora metabolic networks, CarRA quantitative research and the function of CarRA.