論文 - 詳細
| RRC ID | 33461 |
|---|---|
| 著者 | Ahmed A. |
| タイトル | A vector for sequencing long (40-kb) DNA fragments. |
| ジャーナル | Gene |
| Abstract |
An improved vector (pAA113M) has been constructed for sequencing long (40-kb) DNA fragments. The DNA fragment is cloned in the tet gene of the cosmid and subdivided into numerous overlapping segments by IS1-promoted deletions. Plasmids bearing these deletions are fractionated by gel electrophoresis, and shortened further from the opposite end by treatment with specific restriction endonucleases. Thus, a series of short overlapping segments, spread across the entire length of the fragment, become fused to IS1. Each segment can then be sequenced by the dideoxy method using an IS1 primer. The plasmids can replicate either from their normal origin or, in the presence of a filamentous helper phage, from the M13 origin. Hence, each segment can be sequenced as either double-stranded DNA or single-stranded DNA. Sequences of IS1-promoted and restriction enzyme-generated deletions contain overlaps that can be used to assemble the complete 40-kb sequence. |
| 巻・号 | 75(2) |
| ページ | 315-21 |
| 公開日 | 1989-2-20 |
| DOI | 10.1016/0378-1119(89)90277-1 |
| PII | 0378-1119(89)90277-1 |
| PMID | 2541052 |
| MeSH | Base Sequence* Chromosome Deletion Cloning, Molecular DNA / genetics DNA Transposable Elements DNA, Single-Stranded / genetics Escherichia coli / genetics Genetic Techniques Genetic Vectors* Plasmids* Restriction Mapping |
| IF | 2.984 |
| 引用数 | 4 |
| WOS 分野 | GENETICS & HEREDITY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 8 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 原核生物(大腸菌) | pAA113M |