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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
10
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pubmed:dateCreated |
1981-9-15
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pubmed:abstractText |
A method for cloning mRNAs has been used which results in a high yield of recombinants containing complete 5'-terminal mRNA sequences. It is not dependent on self-priming to generate double-stranded DNA and therefore the S1 nuclease digestion step is not required. Instead, the cDNA is dCMP-tailed at its 3'-end with terminal deoxynucleotidyl transferase (TdT). The synthesis of the second strand is primed by oligo(dG) hybridized to the 3'-tail. Double-stranded cDNA is subsequently tailed with dCTP and annealed to dGMP-tailed vector DNA. This approach overcomes the loss of the 5'-terminal mRNA sequences and the problem of artifacts which may be introduced into cloned cDNA sequences. Chicken lysozyme cDNA was cloned into pBR322 by this procedure with a transformation efficiency of 5 x 10(3) recombinant clones per ng of ds-cDNA. Sequence analysis revealed that at least nine out of nineteen randomly isolated plasmids contained the entire 5'-untranslated mRNA sequence. The data strongly support the conclusion that the 5'-untranslated region of the lysozyme mRNA is heterogeneous in length.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-284385,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-325648,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-333389,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-383576,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-390497,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-414220,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-448736,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-455439,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-455453,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-4916553,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-498286,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-514809,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-514812,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-568256,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-593368,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-61066,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-6154933,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-61588,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-62360,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-6251548,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-6934509,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-6993473,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-728434,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-729003,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-7422543,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-7429011,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-768776,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-775445,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-787929,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-802513,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-803499,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6166921-890741
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
May
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pubmed:issn |
0305-1048
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
25
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pubmed:volume |
9
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
2251-66
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:6166921-Avian myeloblastosis virus,
pubmed-meshheading:6166921-Base Sequence,
pubmed-meshheading:6166921-Cloning, Molecular,
pubmed-meshheading:6166921-DNA, Recombinant,
pubmed-meshheading:6166921-Muramidase,
pubmed-meshheading:6166921-Plasmids,
pubmed-meshheading:6166921-RNA, Messenger,
pubmed-meshheading:6166921-RNA-Directed DNA Polymerase
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pubmed:year |
1981
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pubmed:articleTitle |
5'-Terminal sequences of eucaryotic mRNA can be cloned with high efficiency.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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