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Molecular characterization of the Spirometra mansonoides genome: Renaturation kinetics, methylation, and hybridization to human cDNA probes
- Source :
- Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression. 1049:134-144
- Publication Year :
- 1990
- Publisher :
- Elsevier BV, 1990.
-
Abstract
- High molecular weight DNA from pleroceroid larvae of the tapeworm Spirometra mansonoides was purified from isolated nuclei by conventional techniques. The DNA so isolated has a melting temperature (Tm) of 87 degrees C and a guanine plus cytosine (G/C) content of 44%. 5-Methyl cytosine could not be detected in plerocercoid DNA by HPLC analysis of DNA hydrolysates, by radiolabeling 5'-termini of MspI digests with polynucleotide kinase, or by comparing restriction patterns generated by MspI and HpaII. Renaturation kinetics demonstrated that the genome of S. mansonoides contains repetitive as well as single copy sequences and has a genome size estimated at approx. 1.6 X 10(9) bp. Hybridization was carried out between plerocercoid DNA and cDNAs for human beta-actin, alpha-tubulin and growth hormone (hGH). Rationale for this analysis was based on known homologies among actin and tubulin genes in numerous species and on apparent similarities between hGH and a plerocercoid growth factor that may be reflected in similar DNA sequence. Scanning densitometry of dot blots demonstrated that the hGH probe annealed to the same extent at low stringency (1 M NaCl, 55 degrees C) to DNA from plerocercoids, rat liver and chicken erythrocytes; but this interaction was less than to DNA from human lymphocytes, calf thymus and mouse skin. Similar results were obtained when restriction endonuclease digests of these DNAs were analyzed by Southern transfer. Little or no hybridization of the growth hormone probe to plerocercoid DNA was evident at higher stringency (1 M NaCl, 65 degrees C). In contrast, human tubulin and actin probes showed extensive hybridization to pleroceroid restriction fragments under the high stringency conditions.
- Subjects :
- HpaII
Biophysics
Methylation
Biochemistry
Restriction fragment
Nucleic acid thermodynamics
chemistry.chemical_compound
Tubulin
Structural Biology
Sequence Homology, Nucleic Acid
Complementary DNA
Plerocercoid
Genetics
Animals
Spirometra
Repetitive Sequences, Nucleic Acid
Base Composition
biology
Nucleic Acid Hybridization
DNA
Molecular biology
Actins
Blotting, Southern
Restriction enzyme
Genes
chemistry
Growth Hormone
Nucleic Acid Renaturation
biology.protein
Cestoda
Molecular probe
Subjects
Details
- ISSN :
- 01674781
- Volume :
- 1049
- Database :
- OpenAIRE
- Journal :
- Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression
- Accession number :
- edsair.doi.dedup.....512d51ee216769091b1fc024922eacb8