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A simple and rapid nucleotide sequencing strategy and its application in analyzing a rice histone 3 gene
- Source :
- Gene. 45(3)
- Publication Year :
- 1986
-
Abstract
- An improved rapid method for sequencing a target DNA is described. A new plasmid, pAA-PZ1, which contains the origin of replication from phage M13 and a portion of the Tn9 transposon was constructed. A long fragment of target DNA cloned into this vector is progressively shortened in vivo from one end by transposon-mediated deletions. The plasmids carrying different lengths of target DNA are then made into single-stranded DNA in the same host upon infection with an M13 phage and their sequence is determined using the dideoxynucleotide chain-termination method. This method bypasses the in vitro enzymatic manipulations for progressive deletions and requires no subcloning. Using this strategy, we sequenced 1.3 kb of rice DNA containing a histone 3 gene within three weeks.
- Subjects :
- Genetics
Base Sequence
Phagemid
Oryza
General Medicine
DNA
DNA Restriction Enzymes
Biology
Plants
Origin of replication
Sequencing by ligation
Histones
chemistry.chemical_compound
Plasmid
Subcloning
chemistry
Genes
Genomic library
Amino Acid Sequence
Genetic Engineering
In vitro recombination
Plasmids
Subjects
Details
- ISSN :
- 03781119
- Volume :
- 45
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Gene
- Accession number :
- edsair.doi.dedup.....094232b1e2f159ff80a5cdd7d9ae18a6