| Insert |
DNA: cDNA Insert lengths(kb): 3.099999904632568 Tissue: hepatocellular carcinoma Hep G2 cell line Gene product: ligase I, DNA, ATP-dependent(DNA ligase I, ATP-dependent) [LIG1] Target Gene: ligase I, DNA, ATP-dependent |
| Comments |
Restriction digests of the clone give the following sizes (kb): EcoRI--4.6, 1.5, 0.85; HindIII--3.6, 3.3; KpnI--3.7, 3.2; KpnI/PvuI--3.2, 2.6, 1.1; PstI--3.7, 1.8, 1.5; EcoRI/KpnI--3.0, 1.6, 1.5, 0.8. The insert contains the following restriction sites (approximate kb from the 5' KpnI site): EcoRI--0.1, 1.5; PstI--0.75, 2.5; BamHI--0.9; HindIII--2.5. The 1.4 kb EcoRI fragment detects genomic EcoRI fragments of the following approximate sizes (kb): mouse--12.0, (5.5); human--12.0, 8.0, 5.0, 3.1, 2.5. The 1.6 kb EcoRI/KpnI fragment detects genomic EcoRI fragments of the following approximate sizes (kb): mouse--12.0, 5.5, 1.5; human--11.0, 5.5, 3.5, 3.1, 1.5. Complements the cdc9-7 mutation in Saccharomyces cerevisiae. Complements the lig-251 mutation in Escherichia coli. Hybridizes to cosmids f22135, f20031, f22104, and f24839. A full-length cDNA clone. First isolated in the yeast expression vector pAB23BXN (lig2) and subcloned into the present vector as a SalI fragment containing some yeast promoter (GAP) sequence. A KpnI site was introduced into the 5' adaptor such that the full length cDNA can be isolated as a KpnI fragment free of extraneous sequences. The 3.7 kb KpnI fragment includes the vector plus the GAP promoter. The 3.1 kb KpnI fragment is the insert. |
| References |
Barnes DE, et al. Human DNA ligase I cDNA: cloning and functional expression in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 87: 6679-6683, 1990. PubMed: 2204063
Kodama K, et al. In vitro mutagenesis and functional expression in Escherichia coli of a cDNA encoding the catalytic domain of human DNA ligase I. Nucleic Acids Res. 19: 6093-6099, 1991. PubMed: 1956768
Barnes DE, et al. Assignment of the gene encoding DNA ligase I to human chromosome 19q13.2-13.3. Genomics 12: 164-166, 1992. PubMed: 1733856
Deborah E Barnes, personal communication
|
