Abstract：
OBJECTIVE: The Rh blood group system is the most polymorphic human blood group system. Previous studies have investigated variants in the RHD and RHCE promoter. The relevance of these variants to the Chinese Han population is further clarified in this study.
METHODS: In total, 317 donors (223 Rh D-positive [D+], including 20 Del and 94 Rh D-negative [D-]) were randomly selected. The promoter regions and exon 1 of RHD and RHCE were amplified through polymerase chain reaction (PCR) whose products were directly sequenced using forward and reverse primers.
RESULTS: Expected PCR products of the RHD promoter and exon 1 were amplified in 223 D+ individuals, including 20 Del individuals, and were absent in 81 of 94 D- individuals. Expected PCR products of RHCE were observed in all donors. Two single nucleotide variants (SNVs) were observed in the RHD promoter region. Moreover, 11 SNVs were observed in the promoter and exon 1 of RHCE. rs4649082, rs2375313, rs2281179, rs2072933, rs2072932, rs2072931 and rs586178 with strong linkage disequilibria were significantly different between the D+ and D- groups. [A;C] was the most common haplotype in the RHD promoter (NC_000001.11:g.[-1033A>G;-831C>T]). [G;T;T;A;T;A;C;G;A;C;G] was the most predominant haplotype in both total and D- groups. In D+ individuals, [A;C;T;G;C;G;C;G;C;C;C] was the most frequent haplotype in the RHCE promoter (NC_000001.11:g.[-1080A>G;-958C>T;-390T>C;-378G>A;-369C>T;-296G>A;-144C>G;-132G>A;-122C>A;28C>T;48C>G]).
CONCLUSIONS: We speculate that the SNVs/haplotypes found in this article cannot significantly affect gene expression. The present study findings should help elucidate the molecular basis of the polymorphic expression of RHD and RHCE promoter regions.
METHODS: In total, 317 donors (223 Rh D-positive [D+], including 20 Del and 94 Rh D-negative [D-]) were randomly selected. The promoter regions and exon 1 of RHD and RHCE were amplified through polymerase chain reaction (PCR) whose products were directly sequenced using forward and reverse primers.
RESULTS: Expected PCR products of the RHD promoter and exon 1 were amplified in 223 D+ individuals, including 20 Del individuals, and were absent in 81 of 94 D- individuals. Expected PCR products of RHCE were observed in all donors. Two single nucleotide variants (SNVs) were observed in the RHD promoter region. Moreover, 11 SNVs were observed in the promoter and exon 1 of RHCE. rs4649082, rs2375313, rs2281179, rs2072933, rs2072932, rs2072931 and rs586178 with strong linkage disequilibria were significantly different between the D+ and D- groups. [A;C] was the most common haplotype in the RHD promoter (NC_000001.11:g.[-1033A>G;-831C>T]). [G;T;T;A;T;A;C;G;A;C;G] was the most predominant haplotype in both total and D- groups. In D+ individuals, [A;C;T;G;C;G;C;G;C;C;C] was the most frequent haplotype in the RHCE promoter (NC_000001.11:g.[-1080A>G;-958C>T;-390T>C;-378G>A;-369C>T;-296G>A;-144C>G;-132G>A;-122C>A;28C>T;48C>G]).
CONCLUSIONS: We speculate that the SNVs/haplotypes found in this article cannot significantly affect gene expression. The present study findings should help elucidate the molecular basis of the polymorphic expression of RHD and RHCE promoter regions.
摘要：
目的：Rh血型系统是最多态的人类血型系统。先前的研究已经研究了RHD和RHCE启动子中的变体。本研究进一步阐明了这些变异与中国汉族人群的相关性。
方法：总共，317个捐赠者（223个RhD阳性[D+]，随机选择20Del和94RhD阴性[D-])。通过聚合酶链反应（PCR）扩增RHD和RHCE的启动子区域和外显子1，其产物使用正向和反向引物直接测序。
结果：在223个D+个体中扩增了RHD启动子和外显子1的预期PCR产物，包括20名德尔人，94名D人中有81人缺席。在所有供体中观察到RHCE的预期PCR产物。在RHD启动子区域中观察到两个单核苷酸变体(SNV)。此外，在RHCE的启动子和外显子1中观察到11个SNV。rs4649082，rs2375313，rs2281179，rs2072933，rs2072932，rs2072931和rs586178在D组和D-组之间存在显着差异。[A;C]是RHD启动子中最常见的单倍型（NC_000001.11：g。[-1033A>G；-831C>T])。[G;T;T;A;T;A;A;C;G;A;C;G]是总和D组中最主要的单倍型。在D+个人中，[A;C;T;G;C;G;G;G;C;C;C]是RHCE启动子中最常见的单倍型（NC_000001.11：g。[-1080A>G;-958C>T;-390T>C;-378G>A;-369C>T;-296G>A;-144C>G;-132G>A;-122C>A;28C>T;48C>G])。
结论：我们推测本文中发现的SNVs/单倍型不能显着影响基因表达。本研究发现应有助于阐明RHD和RHCE启动子区多态性表达的分子基础。
方法：总共，317个捐赠者（223个RhD阳性[D+]，随机选择20Del和94RhD阴性[D-])。通过聚合酶链反应（PCR）扩增RHD和RHCE的启动子区域和外显子1，其产物使用正向和反向引物直接测序。
结果：在223个D+个体中扩增了RHD启动子和外显子1的预期PCR产物，包括20名德尔人，94名D人中有81人缺席。在所有供体中观察到RHCE的预期PCR产物。在RHD启动子区域中观察到两个单核苷酸变体(SNV)。此外，在RHCE的启动子和外显子1中观察到11个SNV。rs4649082，rs2375313，rs2281179，rs2072933，rs2072932，rs2072931和rs586178在D组和D-组之间存在显着差异。[A;C]是RHD启动子中最常见的单倍型（NC_000001.11：g。[-1033A>G；-831C>T])。[G;T;T;A;T;A;A;C;G;A;C;G]是总和D组中最主要的单倍型。在D+个人中，[A;C;T;G;C;G;G;G;C;C;C]是RHCE启动子中最常见的单倍型（NC_000001.11：g。[-1080A>G;-958C>T;-390T>C;-378G>A;-369C>T;-296G>A;-144C>G;-132G>A;-122C>A;28C>T;48C>G])。
结论：我们推测本文中发现的SNVs/单倍型不能显着影响基因表达。本研究发现应有助于阐明RHD和RHCE启动子区多态性表达的分子基础。
