| Id |
Subject |
Object |
Predicate |
Lexical cue |
| T1 |
0-101 |
Sentence |
denotes |
Use of the Caulobacter crescentus genome sequence to develop a method for systematic genetic mapping. |
| T2 |
102-230 |
Sentence |
denotes |
The functional analysis of sequenced genomes will be facilitated by the development of tools for the rapid mapping of mutations. |
| T3 |
231-423 |
Sentence |
denotes |
We have developed a systematic approach to genetic mapping in Caulobacter crescentus that is based on bacteriophage-mediated transduction of strategically placed antibiotic resistance markers. |
| T4 |
424-571 |
Sentence |
denotes |
The genomic DNA sequence was used to identify sites distributed evenly around the chromosome at which plasmids could be nondisruptively integrated. |
| T5 |
572-687 |
Sentence |
denotes |
DNA fragments from these sites were amplified by PCR and cloned into a kanamycin-resistant (Kan(r)) suicide vector. |
| T6 |
688-787 |
Sentence |
denotes |
Delivery of these plasmids into C. crescentus resulted in integration via homologous recombination. |
| T7 |
788-876 |
Sentence |
denotes |
A set of 41 strains containing Kan(r) markers at 100-kb intervals was thereby generated. |
| T8 |
877-1008 |
Sentence |
denotes |
These strains serve as donors for generalized transduction using bacteriophage phiCr30, which can transduce at least 120 kb of DNA. |
| T9 |
1009-1163 |
Sentence |
denotes |
Transductants are selected with kanamycin and screened for loss of the mutant phenotype to assess linkage between the marker and the site of the mutation. |
| T10 |
1164-1283 |
Sentence |
denotes |
The dependence of cotransduction frequency on sequence distance was evaluated using several markers and mutant strains. |
| T11 |
1284-1435 |
Sentence |
denotes |
With these data as a standard, previously unmapped mutations were readily localized to DNA sequence intervals equivalent to less than 1% of the genome. |
| T12 |
1436-1542 |
Sentence |
denotes |
Candidate genes within the interval were then examined further by subcloning and complementation analysis. |
| T13 |
1543-1666 |
Sentence |
denotes |
Mutations resulting in sensitivity to ampicillin, in nutritional auxotrophies, or temperature-sensitive growth were mapped. |
| T14 |
1667-1816 |
Sentence |
denotes |
This approach to genetic mapping should be applicable to other bacteria with sequenced genomes for which generalized transducing phage are available. |