PMID- 9787074
OWN - NLM
STAT- MEDLINE
DCOM- 19981211
LR  - 20061115
IS  - 0888-7543 (Print)
IS  - 0888-7543 (Linking)
VI  - 53
IP  - 1
DP  - 1998 Oct 1
TI  - Construction of human chromosome 16- and 5-specific circular YAC/BAC libraries by 
      in vivo recombination in yeast (TAR cloning).
PG  - 21-8
AB  - Transformation-associated recombination (TAR) in yeast was exploited for the 
      selective isolation of human DNAs as large circular yeast artificial chromosomes 
      (YACs) from two rodent/human hybrid cell lines containing human chromosomes 5 and 
      16. TAR cloning vectors containing the F-factor origin of replication were 
      constructed for use in these experiments. Presence of the F-factor origin in TAR 
      vectors provides the capability of transferring the YACs generated by in vivo 
      recombination in yeast into Escherichia coli cells and propagating them as 
      bacterial artificial chromosomes (BACs). A high enrichment of human versus rodent 
      YACs was observed during isolation of human DNA from the rodent/human hybrid cell 
      lines. Although <3% of the DNA content in the hybrid cells was human, as many as 
      75% of the transformants contained human YACs. In contrast to the standard YAC 
      cloning method based on in vitro ligation, no human/mouse chimeras were observed 
      during TAR cloning. The constructed human chromosome 16 YAC library had 
      approximately 2.6x coverage, represented by 4320 YAC clones with an average 
      insert size of 80 kb. YAC clones generated from chromosome 16 were successfully 
      converted into BACs by electroporation of DNA isolated from yeast transformants 
      into E. coli. The BAC clones represent approximately 0.6x chromosomal coverage. 
      Pilot YAC and BAC libraries of chromosome 5 have been also constructed. The 
      chromosomal distribution of YAC/BACs from chromosome 5 and chromosome 16 was 
      evaluated by fluorescence in situ hybridization (FISH). The distribution of FISH 
      signals appeared random along the length of each chromosome. We conclude that TAR 
      cloning provides an efficient means for generating representative 
      chromosome-specific YAC/BAC libraries.
CI  - Copyright 1998 Academic Press.
FAU - Kouprina, N
AU  - Kouprina N
AD  - Laboratory of Molecular Genetics, National Institute of Environmental Health 
      Services, Research Triangle Park, North Carolina, 27709, USA. 
      kouprina@niehs.nih.gov
FAU - Campbell, M
AU  - Campbell M
FAU - Graves, J
AU  - Graves J
FAU - Campbell, E
AU  - Campbell E
FAU - Meincke, L
AU  - Meincke L
FAU - Tesmer, J
AU  - Tesmer J
FAU - Grady, D L
AU  - Grady DL
FAU - Doggett, N A
AU  - Doggett NA
FAU - Moyzis, R K
AU  - Moyzis RK
FAU - Deaven, L L
AU  - Deaven LL
FAU - Larionov, V
AU  - Larionov V
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - Genomics
JT  - Genomics
JID - 8800135
RN  - 0 (DNA, Circular)
SB  - IM
MH  - Animals
MH  - Chromosome Mapping
MH  - Chromosomes, Artificial, Yeast/*genetics
MH  - Chromosomes, Human, Pair 16/*genetics
MH  - Chromosomes, Human, Pair 5/*genetics
MH  - Cloning, Molecular/methods
MH  - DNA, Circular/*genetics
MH  - Escherichia coli/genetics
MH  - Gene Library
MH  - Genetic Vectors/genetics
MH  - Humans
MH  - Hybrid Cells
MH  - In Situ Hybridization, Fluorescence
MH  - Recombination, Genetic/*genetics
MH  - Rodentia
MH  - Saccharomyces cerevisiae/genetics
EDAT- 1998/10/27 00:00
MHDA- 1998/10/27 00:01
CRDT- 1998/10/27 00:00
PHST- 1998/10/27 00:00 [pubmed]
PHST- 1998/10/27 00:01 [medline]
PHST- 1998/10/27 00:00 [entrez]
AID - S0888-7543(98)95469-6 [pii]
AID - 10.1006/geno.1998.5469 [doi]
PST - ppublish
SO  - Genomics. 1998 Oct 1;53(1):21-8. doi: 10.1006/geno.1998.5469.