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为了研究1例类孟买型的分子机理,在血清学方法初步鉴定先证者为A类孟买型的基础上,用PCR扩增先证者ABO基因第6、7外显子、FUT1基因(H)和FUT2基因(Se)的编码区序列,PCR产物经割胶纯化后直接测序分析,并应用PCRSSP和基因扫描方法证实测序所发现的突变。FUT1基因突变通过克隆到质粒载体后测序分析。结果表明:直接测序发现先证者ABO血型基因型为A102A102,FUT1基因第682位A→G错义突变、第547-552位两碱基AG缺失(CAGAGAG→CAGAG)突变。克隆证实1条染色体上FUT1基因为A682G突变,导致M228V氨基酸突变;另一条染色体上第547-552位两碱基AG缺失,导致阅读框架发生移码,提前形成终止密码。FUT2基因为分泌基因Se357和弱分泌基因Se357,385杂合(第357位为T/T纯合,385位为A/T杂合)。结论:FUT1基因A682G和547-552delAG双杂合突变是引起该例先证者表现为类孟买型的分子机理,A682G是一个新的引起类孟买型的FUT1基因突变。
In order to study the molecular mechanism of a Mulatta-like type, based on the preliminary identification of probands as serotype A Mumbai type by serological methods, primers 6,7 and 7 of the ABO gene and FUT1 gene (H ) And FUT2 gene (Se). The PCR products were purified by gel excision and sequenced directly. PCRSSP and gene scanning were used to confirm the mutations found by sequencing. FUT1 gene mutation was cloned into plasmid vector and sequenced. The results showed that ABO genotypes of probands were A102A102, A → G at position 682 of FUT1 and CAGAG at position 547-552. The cloning confirmed that the FUT1 gene on the chromosome was A682G mutation, resulting in the mutation of M228V amino acid. The other two bases 547-552 on the other chromosome were deleted, leading to the frameshift of the reading frame and the formation of the stop codon in advance. The FUT2 gene is a hybrid of the secreted gene Se357 and the weakly secreted gene Se357,385 (T / T homozygosity at position 357 and A / T hybrid at position 385). Conclusion: The double heterozygous mutation of FUT1 gene A682G and 547-552delAG are the molecular mechanisms that lead to the case of proband in this case. A682G is a new mutation of FUT1 gene which causes the genus Mumbai.