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Molecular characterization and PCR diagnosis of Thailand deletion of α‐globin gene cluster
Author(s) -
Ko TsangMing,
Tseng LiHui,
Kao ChuanHong,
Lin YuWan,
Hwa HsiaoLin,
Hsu PiMei,
Li ShueFun,
Chuang SouMing
Publication year - 1998
Publication title -
american journal of hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.456
H-Index - 105
eISSN - 1096-8652
pISSN - 0361-8609
DOI - 10.1002/(sici)1096-8652(199802)57:2<124::aid-ajh6>3.0.co;2-y
Subject(s) - breakpoint , genetics , gene , biology , polymerase chain reaction , microbiology and biotechnology , homology (biology) , nucleic acid sequence , mutant , gene cluster , mutation , chromosome
Thailand deletion of α‐Thalassemia (thal) 1 involves the ζ2‐, γζ1‐, α2‐, α1‐, and υ1‐globin genes. In Southeast Asians and Taiwanese, this mutation is the second most common long‐segment deletion of two α‐globin genes, after the Southeast Asian deletion. To define the Thailand deletion breakpoints, we used polymerase chain reaction (PCR) to amplify the normal‐sequence DNA fragments across the breakpoints. The amplified products were sequenced directly or after cloning into pGem®‐3Z or pCR®2.1 vectors. Comparison of the normal and mutant sequences revealed that the 5′ breakpoint lies between nucleotides 1,269 and 1,290 upstream of the initiator codon adenine of the ζ2‐globin gene, and the 3′ breakpoint lies between nucleotides 29,387 and 29,408 downstream of it. A total of 30,677 nucleotides were deleted. Both breakpoints mentioned above lie within the Alu repetitive sequences and an extensive sequence homology is present around the two breakpoints. These findings suggest that homologous recombination is the mechanism by which the deletion occurs. Based on our data, we used three oligonucleotide primers to amplify the regions across the deletion and its corresponding normal sequence. The feasibility of PCR diagnosis was confirmed in 20 carriers with this deletion. Am. J. Hematol. 57:124–130, 1998. © 1998 Wiley‐Liss, Inc.