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培养鼠抗肾综合征出血热病毒衣壳蛋白Ｆ３杂交瘤细胞株，提取总ＲＮＡ，根据鼠源ＩｇＧ抗体基因家族可变区基因碱基序列的特点，设计简并引物，通过逆转录聚合酶链反应，获得抗体轻链可变区和重链可变区基因。分别将其克隆入载体ＰＴ７ＢｌｕｅＴＶｅｃｔｏｒ，选取阳性重组克隆各两个，分别测定了所载重链可变区和轻链可变区基因的碱基序列，比较了不同克隆轻链可变区基因之间和重链可变区基因之间碱基序列的差异；分析了各自的氨基酸框架及其对应蛋白的亲水性。结果显示，两个重链可变区基因碱基序列有４处不同，同源性为９７９％；其中重组克隆ＺＧ３６４５Ｆ所载重链可变区基因有完整的开放阅读框架，对应的蛋白含有丰富的亲水基因，第１１２氨基酸处亲水性最高；另一重组克隆ＺＧ３６４４Ｆ所载重链可变区基因不能通读。两个轻链基因碱基序列有４处不同，同源性为９９１％，重组克隆ＺＧ３６５５Ｆ和ＺＧ３６５７Ｆ所载轻链可变区基因均有完整的开放阅读框架，对应的蛋白均含有丰富的亲水基因，ＺＧ３６５５Ｆ所载基因对应蛋白第６７氨基酸亲水性最高，ＺＧ３６５７Ｆ所载基因对应蛋白第３４氨基酸亲水性最高。
The murine anti-HSV hemagglutinin F3 hybridoma cell line was cultured and the total RNA was extracted. Degenerate primers were designed according to the characteristics of the base sequence of the variable region of the murine IgG antibody gene family. By reverse transcription polymerase chain reaction Reaction to obtain antibody light chain variable region and heavy chain variable region genes. Respectively, cloned into the vector PT7BlueT vector selected positive recombinant clones each two, respectively, the determination of the heavy chain variable region and light chain variable region gene base sequence comparison of different cloned light chain variable region gene The difference between the base sequences of the variable region genes of the heavy chain and the heavy chain was analyzed; and the hydrophilicity of the respective amino acid framework and its corresponding protein was analyzed. The results showed that the two heavy chain variable region gene base sequence has 4 different, homology was 97 9%; wherein the recombinant clone ZG364 5F contained heavy chain variable region genes have a complete open reading frame, the corresponding Protein is rich in hydrophilic genes, the highest hydrophilicity at 112 amino acids; another recombinant cloning ZG364 4F contained heavy chain variable region genes can not read. Two light chain gene base sequence has 4 different, homology was 99.1%, recombinant clones ZG365 5F and ZG365 7F contained in the light chain variable region genes have a complete open reading frame, the corresponding protein Are rich in hydrophilic genes, ZG365 5F contained genes corresponding protein 67 amino acid highest hydrophilic, ZG365 7F contained genes corresponding protein 34 amino acid highest hydrophilicity.