Direct capture and cloning of receptor kinase and peroxidase genes from genomic DNA.
A direct DNA capture and cloning procedure with magnetic bead separation was used to isolate receptor kinase like and peroxidase genes from oat (Avena sativa) and wheat (Triticum aestivum L.) genomic DNA, respectively. In this procedure, the digoxigenin-labeled probe DNA and target genomic DNA fragments were mixed, denatured, and hybridized. The double-helix complexes formed were captured with anti-digoxigenin immunoglobulin-coated magnetic beads and then cloned into either the lambdaBlueSTAR or pUC18 vector. The effectiveness of this procedure was demonstrated by using two specific DNA probes to capture receptor-like kinase genes and surrounding sequences from oat genomic DNA and a peroxidase gene from wheat genomic DNA.
Cheng
Davis W
DW
Department of Biological Sciences, Wayne State University, Detroit, MI 48202, USA.
Armstrong
Ken C
KC
D018554:Avena / Q000201:enzymology / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D015342:DNA Probes; D018744:DNA, Plant / Q000235:genetics* / Q000302:isolation & purification; D017343:Genes, Plant; D005821:Genetic Techniques; D018745:Genome, Plant; D015698:Genomic Library; D010544:Peroxidases / Q000235:genetics*; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*; D014908:Triticum / Q000201:enzymology / Q000235:genetics
D015342:DNA Probes; D018744:DNA, Plant; D010544:Peroxidases; D020794:Receptor Protein-Tyrosine Kinases