DNA-SEQUENCES SPECIFIC FOR CAUCASIAN G3M(B) AND (G) ALLOTYPES - ALLOTYPING AT THE GENOMIC LEVEL
DNA-SEQUENCES SPECIFIC FOR CAUCASIAN G3M(B) AND (G) ALLOTYPES - ALLOTYPING AT THE GENOMIC LEVEL
复制标题
DOI：
10.1007/bf00241259
复制
发表时间：
1994-01-01
期刊：
影响因子：
3.2
通讯作者：
GRUBB, R
中科院分区：
文献类型：
--
作者：
BALBIN, M;GRUBB, A;GRUBB, R
通过对9个高加索人的DNA直接测序，在基因组水平上对G3m(G)和(B)相关氨基酸残基进行了分配。用两个寡核苷酸引物对人IgG3重链基因第二和第三恒定区(C(H)2和C(H)3)的大部分DNA片段进行亚类特异性酶扩增。对血清分型为G3m(B)或G3m(G)纯合子或杂合子G3m(b，g)的个体的扩增DNA序列进行比较，发现氨基酸残基291、296和384的密码子存在差异。这些位置的脯氨酸、苯丙氨酸和丝氨酸对应于G3m(B)，亮氨酸、酪氨酸和天冬氨酸对应于G3m(G)。G3m(b，g)型杂合子个体在这三个位置同时显示G3m(B)和G3m(G)密码子。这三个密码子中的每一个的多态都可以被鉴定为两种限制性内切酶NSP BII和Rsa I的识别位点的出现或丢失。这使得基于聚合酶链式反应(PCR)的分析方法的发展，允许通过分析通过NSP BII和Rsa I消化产生的DNA片段的凝胶迁移率来区分G3m(B)和G3m(G)等位基因。
Assignment of the G3m(g) and (b) correlative amino acid residues was performed at the genomic level by direct sequencing of DNA from nine Caucasian individuals. Two oligonucleotide primers were used for subclass-specific enzymatic amplification of a DNA segment comprising a major portion of the second and third constant region domains (C(H)2 and C(H)3) fo the human IgG3 heavy chain gene. Comparison of the sequences of amplified DNA from individuals serologically typed as homozygous for G3m(b) or G3m(g) or as heterozygous, G3m(b,g), revealed differences in the codons for the amino acid residues 291, 296, and 384. Proline, phenylalanine, and serine at these positions corresponded to G3m(b), and leucine, tyrosine, and asparagine to G3m(g). Heterozygotic individuals, typed G3m(b,g), displayed both the G3m(b) and G3m(g) codons at these three positions. The polymorphism at each of these three codons could be identified either as the appearance, or the loss, of recognition sites for the two restriction endonucleases, Nsp BII and Rsa I. This allowed the development of a polymerase chain reaction (PCR)-based assay permitting the distinction of G3m(b) and G3m(g) alleles by analyzing the electrophoretical mobility of the DNA fragments generated by digestion of the PCR-products with Nsp BII and Rsa I.