MOLECULAR-CLONING AND NUCLEOTIDE-SEQUENCE OF CDNA FOR HUMAN GLUCOSE-6-PHOSPHATE-DEHYDROGENASE VARIANT A(-)
MOLECULAR-CLONING AND NUCLEOTIDE-SEQUENCE OF CDNA FOR HUMAN GLUCOSE-6-PHOSPHATE-DEHYDROGENASE VARIANT A(-)
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DOI：
10.1073/pnas.85.11.3951
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发表时间：
1988-06-01
影响因子：
11.1
通讯作者：
BEUTLER, E
中科院分区：
文献类型：
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作者：
HIRONO, A;BEUTLER, E
葡萄糖-6-磷酸脱氢酶（G6 PD; D-葡萄糖-6-磷酸：NADP+氧化还原酶，EC 1.1.1.49）A（-）是黑人中常见的变体，可导致对药物和感染诱导的溶血性贫血的敏感性。从G6 PDA A（-）男性的EB病毒转化的淋巴母细胞构建cDNA文库。分离的四个cDNA克隆之一含有在其他克隆中未发现的序列，也未在已发表的cDNA序列中发现。该cDNA片段由138个碱基组成，编码46个氨基酸，显然是由内含子7的3“端的选择性剪接产生的。将这些克隆的剩余序列与公开的序列进行比较，发现有三个核苷酸取代：C33 → G，G202 → G。A和A376 →。G.每一个变化都会产生一个新的限制性位点。通过聚合酶链反应扩增来自5个G6 PD A（-）个体的基因组DNA。所有G6 PD A（-）样品中均存在第376位的碱基取代，与G6 PD A（+）中报告的取代相同，并且未检查对照G6 PD B（+）样品。在5个G6 PD A（-）样品中的4个中发现202位的取代，并且没有正常对照样品。在所有的G6 PD A（-）样品和7个G6 PD B（+）对照样品中，在位置33处发现鸟嘌呤，并且推测是在该位置处发现的常见核苷酸。在G6 PD A（-）中发现的与在G6 PD A（+）中发现的相同突变强烈地表明，在个体中出现G6 PD A（-）突变，并添加了导致该酶蛋白体内不稳定性的另一突变。
Glucose-6-phosphate dehydrogenase (G6PD; D-glucose-6-phosphate:NADP+ oxidoreductase, EC 1.1.1.49) A(-) is a common variant in Blacks that causes sensitivity to drug- and infection-induced hemolytic anemia. A cDNA library was constructed from Epstein-Barr virus-transformed lymphoblastoid cells from a male who was G6PDA A(-). One of four cDNA clones isolated contained a sequence not found in the other clones nor in the published cDNA sequence. Consisting of 138 bases and coding 46 amino acids, this segment of cDNA apparently is derived from the alternative splicing involving the 3'' end of intron 7. Comparison of the remaining sequences of these clones with the published sequence revealed three nucleotide substitutions: C33 .fwdarw.G, G202 .fwdarw. A, and A376 .fwdarw. G. Each change produces a new restriction site. Genomic DNA from five G6PD A(-) individuals was amplified by the polymerase chain reaction. The base substitution at position 376, identical to the substitution that has been reported in G6PD A(+), was present in all G6PD A(-) samples and none of the control G6PD B(+) samples examined. The substitution at position 202 was found in four of the five G6PD A(-) samples and no normal control sample. At position 33 guanine was found in all G6PD A(-) samples and seven G6PD B(+) control samples and is, presumable, the usual nucleotide found at this position. The finding of the same mutation in G6PD A(-) as is found in G6PD A(+) strongly suggests that the G6PD A(-) mutation arose in an individual with adding another mutation that causes the in vivo instability of this enzyme protein.