RACE策略克隆鹅抗体轻链基因及分析
loning and Analysis of Ig Light Chain of Goose
目的：在已克隆鹅源抗鹅细小病毒抗体可变区基因的基础上，探索克隆全部抗体轻链基因的成熟方法，为构建单链抗体及体内定位诊断方法研究打基础。方法：从经鹅细小病毒免疫的鹅脾脏，提取脾脏淋巴细胞全RNA，使用SUPERSCRIPT II RT酶和引物GSP-1 对总RNA进行目的基因第一链cDNA的合成，进行3’和5’RACE试验，应用相应的巢式PCR，鉴定后直接与pMD18-T质粒载体连接，转化感受态大肠杆菌DH5α，提重组质粒经PCR鉴定、酶切鉴定后进行序列测定和分析。结果：成功获得的鹅抗体轻链全基因序列符合抗体可变区序列特征。结论：成功克隆抗鹅细小病毒鹅抗体轻链基因序列。
Objective:To clone the Primary structure of the goose’s light chain antibody against goose parvovirus. It provides a basis for further construction of the single chain Fv fragment,which is useful for studing on location diagnose in vivo.METHODS:The V region gene was selected from spleen lymphocyte of the goose immuned by goose parvovirus，extracted of total RNA，reverse transcripted cDNA by SUPERSCRIPT II RT enzyme and primer GSP-1，then amplificated by 3’,5’RACE and cloned into pMD18-T vecter.The recombinant plasmid was identified with restriction endonuclease analysis and PCR，and Sequenced and analyzed. RESULTS:The sequence was in agreement with the characteristics of antibody variable region.Inaddition.CONCLUSION: clone the primary structure of the goose’s light chain antibody against goose parvovirus .The Primary Structure are Proved reasonable and reliable.
王茹丽、刘钧松、李志慧、吕雪峰
分子生物学遗传学生物工程学
鹅抗体轻链测序
GoosentibodyLight chainsequence
王茹丽,刘钧松,李志慧,吕雪峰.RACE策略克隆鹅抗体轻链基因及分析[EB/OL].(2009-12-08)[2025-08-18].http://www.paper.edu.cn/releasepaper/content/200912-240.点此复制
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