Cloning and sequencing of KIR2DL1 framework gene cDNA and identification of a novel allele / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
; (6): 694-697, 2016.
Article
en Zh
| WPRIM
| ID: wpr-345377
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To develop an assay for cDNA cloning and haplotype sequencing of KIR2DL1 framework gene and determine the genotype of an ethnic Han from southern China.</p><p><b>METHODS</b>Total RNA was isolated from peripheral blood sample, and complementary DNA (cDNA) transcript was synthesized by RT-PCR. The entire coding sequence of the KIR2DL1 framework gene was amplified with a pair of KIR2DL1-specific PCR primers. The PCR products with a length of approximately 1.2 kb were then subjected to cloning and haplotype sequencing.</p><p><b>RESULTS</b>A specific target fragment of the KIR2DL1 framework gene was obtained. Following allele separation, a wild-type KIR2DL1*00302 allele and a novel variant allele, KIR2DL1*031, were identified. Sequence alignment with KIR2DL1 alleles from the IPD-KIR Database showed that the novel allele KIR2DL1*031 has differed from the closest allele KIR2DL1*00302 by a non-synonymous mutation at CDS nt 188A>G (codon 42 GAG>GGG) in exon 4, which has caused an amino acid change Glu42Gly. The sequence of the novel allele KIR2DL1*031 was submitted to GenBank under the accession number KP025960 and to the IPD-KIR Database under the submission number IWS40001982. A name KIR2DL1*031 has been officially assigned by the World Health Organization (WHO) Nomenclature Committee.</p><p><b>CONCLUSION</b>An assay for cDNA cloning and haplotype sequencing of KIR2DL1 has been established, which has a broad applications in KIR studies at allelic level.</p>
Texto completo:
1
Índice:
WPRIM
Asunto principal:
Haplotipos
/
Secuencia de Bases
/
Química
/
China
/
Clonación Molecular
/
Análisis de Secuencia de ADN
/
ADN Complementario
/
Mutación Missense
/
Alelos
/
Receptores KIR2DL1
Tipo de estudio:
Diagnostic_studies
Límite:
Humans
/
Male
País/Región como asunto:
Asia
Idioma:
Zh
Revista:
Chinese Journal of Medical Genetics
Año:
2016
Tipo del documento:
Article