PMID- 1472941
OWN - NLM
STAT- MEDLINE
DCOM- 19930129
LR  - 20191028
IS  - 1054-9803 (Print)
IS  - 1054-9803 (Linking)
VI  - 1
IP  - 3
DP  - 1992 Feb
TI  - A method for specific amplification and PCR sequencing of individual members of 
      multigene families: application to the study of steroid 21-hydroxylase 
      deficiency.
PG  - 181-6
AB  - Mutations at the human HLA-linked CYP21B locus are responsible for 21-hydroxylase 
      deficiency, a recessively inherited disorder of steroidogenesis. The scope for 
      PCR-based analysis of the CYP21B gene has been restricted by the very high 
      sequence homology between CYP21B and a closely related pseudogene, CYP21A. Here 
      we describe a novel PCR sequencing strategy that allows the independent 
      amplification of the entire CYP21B coding sequence and the subsequent 
      enzyme-mediated conversion of the PCR product to a single-stranded form for 
      dideoxy sequencing. We have used this approach to characterize the 21-hydroxylase 
      deficiency allele associated with HLA-B55, the most frequent HLA marker 
      associated with a CYP21B point mutation in the British population, and also an 
      HLA-B35 associated allele of Asian origin. Allele-specific oligonucleotide (ASO) 
      hybridization analyses have confirmed the selective amplification of CYP21B genes 
      and the identity of the pathological mutations. The method can be adapted to 
      permit selective amplification and PCR sequencing of individual closely related 
      members of other multigene families and small-copy-number repetitive DNA 
      families.
FAU - Collier, S
AU  - Collier S
AD  - University Department of Medical Genetics, St. Mary's Hospital, Manchester, UK.
FAU - Tassabehji, M
AU  - Tassabehji M
FAU - Strachan, T
AU  - Strachan T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - PCR Methods Appl
JT  - PCR methods and applications
JID - 9201445
RN  - 0 (HLA Antigens)
RN  - EC 1.14.14.16 (Steroid 21-Hydroxylase)
SB  - IM
GS  - CYP21B
MH  - Adrenal Hyperplasia, Congenital
MH  - Base Sequence
MH  - DNA Mutational Analysis
MH  - Evaluation Studies as Topic
MH  - Genetic Linkage
MH  - HLA Antigens/genetics
MH  - Humans
MH  - Molecular Sequence Data
MH  - *Multigene Family
MH  - Nucleic Acid Amplification Techniques
MH  - Polymerase Chain Reaction/*methods
MH  - Steroid 21-Hydroxylase/*genetics
EDAT- 1992/02/01 00:00
MHDA- 1992/02/01 00:01
CRDT- 1992/02/01 00:00
PHST- 1992/02/01 00:00 [pubmed]
PHST- 1992/02/01 00:01 [medline]
PHST- 1992/02/01 00:00 [entrez]
AID - 10.1101/gr.1.3.181 [doi]
PST - ppublish
SO  - PCR Methods Appl. 1992 Feb;1(3):181-6. doi: 10.1101/gr.1.3.181.