PMID- 7603471
OWN - NLM
STAT- MEDLINE
DCOM- 19950810
LR  - 20191031
IS  - 0890-8508 (Print)
IS  - 0890-8508 (Linking)
VI  - 9
IP  - 2
DP  - 1995 Apr
TI  - New members of the 3 beta-hydroxysteroid dehydrogenase gene family.
PG  - 121-8
AB  - Several bands of hydridization are detected when southern blots of human genomic 
      DNA are proved with cDNA of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) type 
      I. Two experimental approaches were adopted to estimate the size of the 3 
      beta-HSD gene family. Firstly, primer designed to amplify 3 beta-HSD type I and 
      II genes were found on occasion to amplify DNA products of appropriate length but 
      which were resolved as distinct sequences by denaturing gradient gel 
      electrophoresis (DGGE). Five of these novel bands were cloned and their sequences 
      were found to be closely related to 3 beta-HSD types I and II. Secondly, 57 
      genomic clones were selected from two lambda genomic libraries by hybridization 
      with exonic probes of 3 beta -HSD type I. These were screened for novel members 
      of the gene family by pcr amplification using various combinations of PCR primers 
      to the type I and II genes, particularly those primers that previously amplified 
      novel PCR products from genomic DNA. Amplification products from (lambda) clones 
      were screened for novel sequences by DGGE. As a result of these approaches, at 
      least five new members of the 3 beta-HSD gene family were found, one of which 
      locates to the 3 beta -HSD type I and II gene cluster on 1p13. The existence of 
      additional closely related but distinct members of the gene family should be 
      recognized as a potential complication when screening PCR fragments for mutations 
      in the type I and II genes. DGGE was found to be an exceedingly rapid means of 
      screening amplification products from (lambda) clones to search for novel members 
      of the gene family.
FAU - McBride, M W
AU  - McBride MW
AD  - Institute of Genetics, Glasgow University.
FAU - Russell, A J
AU  - Russell AJ
FAU - Vass, K
AU  - Vass K
FAU - Forster, V
AU  - Forster V
FAU - Burridge, S M
AU  - Burridge SM
FAU - Morrison, N
AU  - Morrison N
FAU - Boyd, E
AU  - Boyd E
FAU - Ponder, B A
AU  - Ponder BA
FAU - Sutcliffe, R G
AU  - Sutcliffe RG
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Mol Cell Probes
JT  - Molecular and cellular probes
JID - 8709751
RN  - 0 (DNA Primers)
RN  - 0 (DNA Probes)
RN  - 0 (Isoenzymes)
RN  - 0 (Recombinant Proteins)
RN  - EC 1.1.- (3-Hydroxysteroid Dehydrogenases)
SB  - IM
MH  - 3-Hydroxysteroid Dehydrogenases/biosynthesis/chemistry/*genetics
MH  - Amino Acid Sequence
MH  - Base Sequence
MH  - Blotting, Southern
MH  - Cloning, Molecular
MH  - DNA Primers
MH  - DNA Probes
MH  - Exons
MH  - Genomic Library
MH  - Humans
MH  - Isoenzymes/biosynthesis/chemistry/genetics
MH  - Molecular Sequence Data
MH  - *Multigene Family
MH  - Polymerase Chain Reaction
MH  - Recombinant Proteins/biosynthesis/chemistry
MH  - Sequence Homology, Amino Acid
EDAT- 1995/04/01 00:00
MHDA- 1995/04/01 00:01
CRDT- 1995/04/01 00:00
PHST- 1995/04/01 00:00 [pubmed]
PHST- 1995/04/01 00:01 [medline]
PHST- 1995/04/01 00:00 [entrez]
AID - 10.1016/s0890-8508(95)80036-0 [doi]
PST - ppublish
SO  - Mol Cell Probes. 1995 Apr;9(2):121-8. doi: 10.1016/s0890-8508(95)80036-0.