[35] Cross-species polymerase chain reaction: Cloning of TATA box-binding proteins
This chapter discusses an application of the polymerase chain reaction (PCR) to isolate the genomic or complementary DNA (cDNA) fragments of a gene from one species, given that the polypeptide or DNA sequence of the gene from a second species is known. The PCR technique is extremely sensitive, which...
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| Veröffentlicht in: | Methods in Enzymology 1993, Vol.218, p.493-507 |
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| creator | Peterson, Michael Gregory Dian, Robert |
| description | This chapter discusses an application of the polymerase chain reaction (PCR) to isolate the genomic or complementary DNA (cDNA) fragments of a gene from one species, given that the polypeptide or DNA sequence of the gene from a second species is known. The PCR technique is extremely sensitive, which means that the utmost care must be taken to avoid cross-contamination of samples, especially from micropipetting equipment. Physically separating work areas and equipment used for setting up the reactions and the subsequent analysis of the PCR products is perhaps the best way to avoid cross-contamination. The complete sequence of Drosophila and human t-butylbicyclophosphorothionates (TBPs) is analyzed in this chapter. This analysis shows that of the 10 regions of the yeast amino acid sequence chosen for designing PCR primers, three show perfect identities between the two species and other three show one amino acid sequence difference between the two species. |
| doi_str_mv | 10.1016/0076-6879(93)18037-D |
| format | Article |
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| ispartof | Methods in Enzymology, 1993, Vol.218, p.493-507 |
| issn | 0076-6879 1557-7988 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_75779153 |
| source | MEDLINE; Elsevier ScienceDirect Journals; ScienceDirect eBooks |
| subjects | Amino Acid Sequence Animals Base Sequence Cloning, Molecular - methods DNA DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism DNA-Directed DNA Polymerase Drosophila melanogaster - genetics Drosophila melanogaster - metabolism Escherichia coli - genetics HeLa Cells Humans Indicators and Reagents Molecular Sequence Data Oligodeoxyribonucleotides Polymerase Chain Reaction - methods Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Species Specificity Taq Polymerase TATA Box TATA-Box Binding Protein Transcription Factors - genetics Transcription Factors - metabolism Transcription Factors, TFIII |
| title | [35] Cross-species polymerase chain reaction: Cloning of TATA box-binding proteins |
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