SBIR-STTR Award

Site Specific Cloning Of Genomic DNA In Vivo
Award last edited on: 11/21/05

Sponsored Program
SBIR
Awarding Agency
NIH : NICHD
Total Award Amount
$99,996
Award Phase
1
Solicitation Topic Code
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Principal Investigator
Kevin L Bentley

Company Information

Genaissance Pharmaceuticals Inc (AKA: BIO Pharmaceuticals Inc)

5 Science Park
New Haven, CT 06511
   (203) 773-1450
   contact-us@genaissance.com
   www.genaissance.com
Location: Multiple
Congr. District: 03
County: New Haven

Phase I

Contract Number: 1R43HD035417-01
Start Date: 00/00/00    Completed: 00/00/00
Phase I year
1997
Phase I Amount
$99,996
The mechanism of gap repair in yeast in the basis for development of a facile method for in vivo cloning of large fragments of genomic DNA. A yeast-bacteria shuttle vector, pCLASPER, linearized to expose interchangeable site-specific recombinogenic ends, can target homologous sequences in co-transformed genomic DNA. The genomic region delimited by the recombinogenic ends is recovered in pCLASPER via the high frequency of yeast homologous recombination. Recombinants are identified by acquisition of a yeast selectable marker. The cloned products can be transferred to E. Coli for large-scale growth and plasmid DNA extraction by standard alkaline lysis methods. Methods will be devised and adapted for optimal co-transformation efficiency of linearized vector and uncloned genomic DNA in S. cerevisiae, the length of homology necessary for target-specific cloning will be determined, and sources for generating targeting sequence will be tested. Targeted in vivo cloning provides a method for selective recovery of a single, specific genomic fragment from one or many individuals of a population, without library construction and of a size larger than amplified by long-range PCR. Applications include sequence gap closure , mapping and subcloning, haplotyping, population sampling mutational analysis, gene funtion, and phylogientic comparisons.

Thesaurus Terms:
biotechnology, gene targeting, genome, molecular cloning, transfection vector bacterial genetics, cell transformation, fungal genetics, method development, nucleic acid sequence, transposon /insertion element Escherichia coli, Saccharomyces cerevisiae, polymerase chain reaction, site directed mutagenesisNational Institute of Child and Human Development (NICHD)

Phase II

Contract Number: ----------
Start Date: 00/00/00    Completed: 00/00/00
Phase II year
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Phase II Amount
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