新生儿免疫球蛋白重链可变区基因多样性分析

Analysis of the Immunoglobulin Heavy Chain Variable Region gene Fingerprinting of Human Neonates

目的 构建不同胎龄新生儿IgH基因指纹图谱 ,探讨新生儿成熟度对IgH基因谱型多样性的影响 .方法提取 2 7～ 42周新生儿RNA、进行RT -PCR反应、6%变性聚丙烯酰胺凝胶电泳和银染显色 ,获得不同胎龄新生儿IgH基因指纹图谱 ,用Tiger凝胶图像分析仪进行曲线转化和分析 .结果 各家族指纹图谱转化后的曲线下面积积分值 ,在 2 8～ 3 2周、3 3～ 3 6周和 3 7～ 42周新生儿之间无差异 ;但胎龄 2 7周新生儿曲线下面积小于胎龄 2 8～ 42周新生儿 .结论 2 8周可能是人类IgH基因谱型发育由不完善到趋于完善的一个转折点 ,2

Objective To develop IgH gene fingerprinting of human neonates with different gestational age (GA), as well as study the effect of neonatal maturity on IgH repertoire. Methods Peripheral blood from the neonates with GA of 27～42 weeks were collected. RNA was extracted from mononuclear cells. RT-PCR was used to amplify the IgH gene, and PCR products were separated by electrophoresis and stained on 6% denaturing PAGE gel. Results The differences of area below the curve in neonates with GA 28～32、33～36 and 37～42 weeks were not significant, but those in neonates with GA 27 weeks were obviously less. Conclusions 28th week of GA may be a turning point in course of IgH gene repertoire developing. To some extent, fingerprintings of neonates with GA from 28 to 42 weeks apporach to be stable.