Clone mapping

Jiang W, WW Metcalf, K-S Lee, BL Wanner (1995) Molecular cloning, mapping, and regulation of the pho regulon genes for phosphonate breakdown by the phosphonatase pathway of Salmonella typhimurium LT2. J Bacteriol 111 6411-6421. 

D. T. Bonthron. 1-Asparaginase tl of Escherichia coli K-12 cloning, mapping and... 

Rice, C. M., Levis, R., Strauss, J. H., and Huang, H. V. (1987). Production of infectious RNA transcripts from Sindbis virus cDNA clones Mapping of lethal mutations, rescue of a temperature sensitive marker, and in vitro mutagenesis to generate defined mutants./. Virol. 61, 3809-3819. 

Gebe, J.A., Kiener, P.A., Ring, H.Z., Li, X., Francke, U., and Amffo, A., 1997, Molecular cloning, mapping to human chromosome 1 q21-q23, and cell binding characteristics of Spalpha, a new member of the scavenger receptor cysteine-rich (SRCR) family of proteins./. Biol. Chem. 272 6151-6158. 

Workers in the early 1970s recognized that restriction enzymes provided tools not only for DNA mapping but also for constmction of new DNA species not found in nature. A collection of recombinant DNA species consisting of many passenger sequences joined to identical vector molecules is called a hbrary. Individual recombinant DNAs are isolated from single clones of the Hbrary for detailed analysis and manipulation. 

Eig. 5. Restriction map of the yeast artificial chromosome (YAC) vector used for cloning very large fragments of eukaryotic DNA. Terms defined in text... 

After a desired clone is obtained and mapped with restriction enzymes, further analysis usually depends on the deterrnination of its nucleotide sequence. The nucleotide sequence of a new gene often provides clues to its function and the stmcture of the gene product. Additionally, the DNA sequence of a gene provides a guidepost for further manipulation of the sequence, for example, lea ding to the production of a recombinant protein in bacteria. 

A number of the genes involved in the biosynthesis of thiamine in E. coli (89—92), i hium meliloti (93), B. suhtilis (94), and Schi saccharomycespomhe (95,96) have been mapped, cloned, sequenced, and associated with biosynthetic functions. Thiamine biosynthesis is tightly controlled by feedback and repression mechanisms limiting overproduction (97,98). A cost-effective bioprocess for production of thiamine will require significant additional progress. 

RONEN G, CARMEL-GOREN L, ZAMIR D and HIRSCHBERG J (2000) An alternative pathway to 13-carotene formation in plant chromoplast discovered by map-based cloning of beta and old-gold color mutations in tomato , Proc Natl Acad Sci, 97, 11102-7. 

In maize, many phenotypic mutants have been associated with cloned genes by a combination of HPLC analysis of specific intermediate metabolite accumulation, RT-PCR and immunolocalization of candidate genes, and recombinant inbred mapping of candidate cDNAs. Psyl was cloned by transposon tagging and later shown to be functional in the color complementation system and to be the specific... 

DNA restriction analysis of the corresponding cosmids revealed one common c oRJ fragment of 9.2 kbp. The restriction map of the DNA region covered by the overlapping clones is shown in Fig 1. 

The XE6 DNA was digested with several restriction enzymes and analysed via Southern blots using a probe derived fi-om the pgaW gene. A restriction map of the X.E6 clone revealed that the complete gene should be present on a 3.0 kb EcoRl fi agment (see Fig. 1). 

Figure 1. The detailed restriction map of the 3.0 kb EcdFl fragment of the XE6 clone. The position of pgaE gene is shown by the black bar. This fragment was cloned into pBluescript SK and further analysed.

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