Linked Life Data

3026927

Source:http://linkedlifedata.com/resource/pubmed/id/3026927

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1987-3-19
pubmed:databankReference
pubmed:abstractText
We wished to determine whether there is any specific sequence downstream of the start point of the SP6 promoter which is required for its function in the plasmid pSP64 (Melton et al., 1984). Lack of such specificity would permit in vitro synthesis of an RNA molecule having a 5'-terminal sequence identical to its wild-type in vivo counterpart. To test its requirement, we replaced all of the SP6 sequence downstream of the transcription start point with heterologous nucleotides (nt) and found that any sequence will suffice to permit efficient and accurate transcription. These results permitted construction of plasmids for synthesis of 'authentic' transcripts from cloned DNA. In one case, by an oligodeoxynucleotide-mediated site-specific deletion, we placed the start point of yeast gene TCM1 at nt + 2 of the SP6 promoter and produced in vitro TCM1 mRNA with a wild-type 5'-terminal sequence. We also constructed a vector, pSP64 delta 1, in which the SalI/AccI/HincII recognition sites of pSP64 reside at nt + 2 through + 7. Plasmids such as pSP64 delta 1 may be more useful in some cases as insertion of any DNA fragment at one of these three sites will yield a transcript in which only two to four nt are derived from the vector.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
0378-1119
pubmed:author
pubmed:issnType
Print
pubmed:volume
46
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
57-64
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1986
pubmed:articleTitle
Plasmids allowing transcription of cloned DNA by Salmonella typhimurium phage SP6 RNA polymerase to produce RNAs with authentic 5'-terminal sequences.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, Non-P.H.S.