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Cloning Experiments (cloning + experiment)
Selected AbstractsB-F DNA sequence variability in Brazilian (blue-egg Caipira) chickensANIMAL GENETICS, Issue 4 2004C. A. V. Lima-Rosa Summary A total of 100 chickens from the Brazilian (blue-egg Caipira) native breed were studied in relation to exon 2 of the B-F genes of the major histocompatibility complex (MHC) region. After a first screening on 100 birds, 22 animals were selected for amplification, cloning and sequencing experiments of exons 2,4 (a total of 1048 bp) of their DNA. Twenty-three sequences were obtained, of which at least 10 appear novel. Inferred protein sequences were compared with those previously described, totalling 41 different sequences with amino acid changes in 33 of the 88 sites in ,1, and 34 of the 91 sites in ,2 domains. Allele expression was investigated in these animals through cloning experiments. The blue-egg Caipira chickens may provide a source of novel B-F alleles for genetic improvement programmes. [source] Stability of recombinant plasmids on the continuous culture of Bifidobacterium animalis ATCC 27536BIOTECHNOLOGY & BIOENGINEERING, Issue 2 2003Antonio González Vara Abstract Bifidobacterium animalis ATCC 27536 represents among bifidobacteria a host-model for cloning experiments. The segregational and structural stabilities of a family of cloning vectors with different molecular weights but sharing a common core were studied in continuous fermentation of the hosting B. animalis without selective pressure. The rate of plasmid loss (R) and the specific growth rate difference (,,) between plasmid-free and plasmid-carrying cells were calculated for each plasmid and their relationship with plasmid size was studied. It was observed that both R and the numerical value of ,, increased exponentially with plasmid size. The exponential functions correlating the specific growth rate difference and the rate of plasmid loss with the plasmid molecular weight were determined. Furthermore, the smallest of the plasmids studied, pLAV (4.3-kb) was thoroughly characterized by means of its complete nucleotide sequence. It was found that it contained an extra DNA fragment, the first bifidobacterial insertion sequence characterised, named IS 1999. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 84: 145,150, 2003. [source] Large scale purification of linear plasmid DNA for efficient high throughput cloningBIOTECHNOLOGY JOURNAL, Issue 9 2010Dr. Marjolaine Noirclerc-Savoye Abstract In this report we describe a rapid, simple, and efficient method for large-scale purification of linear plasmid DNA to answer demand from high-throughput gene cloning. The process is based on the separation of the linear vector from small DNA fragments by anion exchange chromatography. Gene cloning experiments by restriction/ligation or the In-Fusion(tm) technique confirmed the high quality of the linearized vector as 100% of the genes were successfully cloned. [source] Cover Picture: Chemical Restriction: Strand Cleavage by Ammonia Treatment at 8-Oxoguanine Yields Biologically Active DNA (ChemBioChem 7/2003)CHEMBIOCHEM, Issue 7 2003Christoph Meyer Dr. Abstract The cover picture shows a chemical restriction method in which DNA double strands are cleaved at the position of the artificial base 8-oxoguanine (Goxo). As as result of the mild cleavage conditions, the DNA fragments remain biologically active and can be used in gene cloning experiments. As an example, the lac Z, gene was amplified by PCR with 8-oxoguanine-modified primers, restricted by treatment with ammonia, ligated into a plasmid vector, transformed in Escherichia coli cells, and screened for blue colonies. This method guarantees efficiencies comparable to the standard cloning procedure, and allows the design of any 3, overhang, independent of the sequence of the cloned DNA. Further information can be found in the article by Giese et. al. on pp. 610,614 [source] | ||||||||||