PMID- 21080280
OWN - NLM
STAT- MEDLINE
DCOM- 20110303
LR  - 20211020
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Print)
IS  - 1064-3745 (Linking)
VI  - 693
DP  - 2011
TI  - Engineering BAC reporter gene constructs for mouse transgenesis.
PG  - 163-79
LID - 10.1007/978-1-60761-974-1_10 [doi]
AB  - A culmination of large-scale ideas and efforts has truly allowed for the use of 
      large genomic DNA clones housed in Bacterial Artificial Chromosome (BAC) vectors 
      for biological research. Fundamental advances that have allowed this to happen 
      include (1) the completion of genome sequencing projects and the establishment of 
      highly annotated web-accessible databases allowing for the rapid identity and 
      purchase of BAC clones containing genes of interest. (2) The generation of 
      methodologies to modify BACs genetically, allowing for the rapid creation of gene 
      targeting constructs or transgenic reporter gene constructs using homologous 
      recombination in bacteria.Recent efforts on our part have capitalized on these 
      advances by using BACs and bacterial recombination methods to generate 
      fluorescent protein reporter transgenic mice to study skeletal biology. The 
      rationale for using BAC genomic DNA clones to engineer reporter gene constructs 
      is based on their much larger size, thus increasing the likelihood that most, if 
      not all, of a gene's respective cis regulator elements are present, giving a 
      truer representation of the endogenous gene's expression. In a relatively short 
      amount of time, we have become extremely proficient at generating BAC reporters. 
      Contrary to the widely perceived notion that working with BACs is complex and 
      difficult, we decided to write this chapter to encourage laboratories that are 
      currently using traditional molecular cloning methods to engineer transgenic DNA 
      constructs to strongly consider learning BAC methodologies. As an example, we 
      walk through the steps we took to generate the transgenic reporter mouse line, 
      Tenascin C (TNC)-mCherry.
FAU - Fu, Yu
AU  - Fu Y
AD  - Department of Reconstructive Sciences, School of Dental Medicine, University of 
      Connecticut Health Center, Farmington, CT, USA.
FAU - Maye, Peter
AU  - Maye P
LA  - eng
GR  - R21 RR021707/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Chromosomes, Artificial, Bacterial/*genetics
MH  - *Gene Transfer Techniques
MH  - Genes, Reporter/*genetics
MH  - Green Fluorescent Proteins/genetics/metabolism
MH  - Mice
MH  - Mice, Transgenic/*genetics
MH  - Recombination, Genetic/genetics
PMC - PMC5647146
MID - NIHMS909940
EDAT- 2010/11/17 06:00
MHDA- 2011/03/04 06:00
PMCR- 2017/10/18
CRDT- 2010/11/17 06:00
PHST- 2010/11/17 06:00 [entrez]
PHST- 2010/11/17 06:00 [pubmed]
PHST- 2011/03/04 06:00 [medline]
PHST- 2017/10/18 00:00 [pmc-release]
AID - 10.1007/978-1-60761-974-1_10 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2011;693:163-79. doi: 10.1007/978-1-60761-974-1_10.