| 猪圆环病毒1型ORF2基因的克隆与序列分析 |
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| 作者姓名： | 靳玉芬 郝贵增 何宏轩 |
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| 作者单位： | 郑州牧专动物医药系 郑州450011(徐耀辉，乔宏兴，臧金灿，卢建洲)，河南省商丘市动物检疫站 商丘476000(李翠平) |
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| 基金项目： | 河南省科技攻关重点项目＂畜禽链球菌病、沙门氏菌病快速诊断及防治关键技术研究与应用（072102130009）＂资助. |
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| 摘 要： | 【研究目的】提取沙门氏菌鞭毛蛋白并鉴定其抗原性;【方法】选用鸭沙门氏菌经半固体培养基诱导鞭毛,经肉汤培养后,用酸裂解、经差速离心、硫酸铵沉淀,分离出粗制鞭毛蛋白。将粗制的鞭毛蛋白用SDS-PAGE和磷钨酸负染投射电镜观察,并包板做ELISA检测其抗原性;【结果】SDS-PAGE显示提取的鞭毛蛋白相对分子质量约为58.0KD,且纯度较高,抗原性较好,每1000ml培养液可获得32.8mg鞭毛蛋白;【结论】酸裂解法可获得高质量的鞭毛蛋白,抗原性较好,为禽副伤寒沙门氏菌病诊断方法的建立奠定了基础。
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| 关 键 词： | 鸭沙门氏菌 鞭毛 提取 抗原性鉴定 |
Cloning and Sequencing Analysis of ORF2 Gene of Porcine Circovirus 1 |
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| Authors: | Jin Yufen Hao Guizeng He Hongxuan |
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| Institution: | Department of Veterinary Medicine, Zhengzhou College of A nimal Husbandry Engineering, Zhengzhou 450011; 2Henan, Shangqiu Animal Quarantine Station, Shangqiu 476000 |
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| Abstract: | Objective]To separate and identify the flagellin from Salmonella anatum.Method]Salmonella anatum was induced with semisolid medium for flagellin,and cultured with M broth,then obtained by centrifuging.The flagellin was separated by acid-splitting and differential centrifugation,ammonium sulate precipitationi.The obtained flagellin was evaluated by sodiummdodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE),phosphotungstic acid negative staining and electron microscopy.Antigencity of flagellin was detected by direct ELISA.Results]SDS-PAGE showed that the purified flagellin protein molecular weight was 58KD;the protein possessed good antigencity and 1000 milliliters of culture produced 32.8mg flagellin.Conclusion]The flagellin with good antigencity from Salmonella anatum can be obtained by acid-splitting.The purified flagellin may lay the foundation for developping methods for avian paratyphoid diagnosis. |
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| Keywords: | Salmonella anatum Flagellin Separation Antigencity identification |
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