Molecular basis of ?-thalassemia in Thailand: analysis of ?-thalassemia mutations using the polymerase chain reaction

Abstract

beta-Thalassemia mutations in 71 chromosomes of Thai patients from the northeast, the middle and the south of the country were investigated using dot blot hybridization of PCR (polymerase chain reaction)-amplified DNA with allele-specific oligonucleotide probes. Eight different known molecular defects were detected, at different frequencies. There was an amber mutation in codon 17, a C-T transversion at position 654 of IVS-2, a frameshift mutation between codons 71 and 72, an A-G transition at nucleotide -28 within the TATA box (known as Chinese mutations), a G-T transversion at position 1 of IVS-1 (an Indian mutation), a 4 bp deletion in codons 41/42 and a G-C transversion at position 5 of IVS-1 (described as both Chinese and Indian mutations) and a Thai original mutation, an ochre mutation in codon 35. Analysis of the three unknown alleles by DNA sequencing of the cloned DNA fragment amplified by PCR revealed an A-G substitution at the second position of the codon for amino acid 19 (AAC-AGC). The analytic approach used in the present study and the characteristic distribution of mutations in each region of Thailand will prove useful for setting up a prenatal diagnosis program.

Mutations and polymorphisms of Hirschsprung disease candidate genes in Thai patients

“…Geographically, the region is situated in the middle part of the Malayu peninsula, between the mainland of Thailand and Malaysia. Previous study has suggested that the population in this region was an admixture of two ethnic factions, the Chinese-Thai extract from the Thai mainland and the Malay-Thai extract, which is predominantly Muslim (Fucharoen et al 1989;Laosombat et al 1992). …”

Coexistence of Southeast Asian ovalocytosis and β‐thalassemia: A molecular and hematological analysis

“…Hb analysis using an automated cation exchange HPLC (VARIANT, Bio-Rad Laboratories, Hercules, CA) demonstrated Hb A 2 A pattern with the amount of Hb A 2 at 5.6%, characteristic of a b-thalassemia carrier. Further DNA analysis of the b-globin gene by the polymerase chain reaction (PCR) routinely run in our laboratory revealed that the proband carried a -28 A-G/b + -thalassemia mutation which is one of the common b-thalassemia genes in Thailand [7]. DNA analysis of a-globin gene cluster identified no a-thalassemia mutations, namely, a…”

PI * S iiyama , a deficiency gene of alpha1-antitrypsin: Evidence for the occurrence in western Japan

“…Amplification of the AAT gene of the proband and direct sequencing of the PCR-amplified products were performed under the condition as described (Fucharoen et al, 1989;Matsunaga et al, 1990). The sequence of primers used for amplification of the coding exons of the AAT gene was as follows :…”

“…Amplified products of exon II were dot-blotted onto nylon membrane and hybridized with allele-specific oligonucleotide probes at 42~ for 1 hr and then washed at 55~ for l0 rain (Fucharoen et al, 1989). The sequence of the two probes was as follows:…”