Linked Life Data

12135880

Source:http://linkedlifedata.com/resource/pubmed/id/12135880

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2002-7-23
pubmed:abstractText
Successful embryonic development is dependent on time and location-specific expression of appropriate genes. Unfortunately, information on stage-specific gene expression during early embryonic development in the bovine is lacking. In the present study, we compared gene expression between in vitro-produced Day 7-8 intact blastocysts (driver) and Day 9-10 hatched blastocysts (tester) using suppression-subtractive hybridization. Pools of 30 embryos for both driver and tester were used in the RNA extraction process. From limited amounts of starting material ( approximately 400 ng of total RNA), a reverse transcription-polymerase chain reaction (PCR) procedure was used to amplify the mRNA and generate sufficient cDNA to conduct suppression-subtractive hybridization. The subtracted cDNA products were cloned, and 126 cDNAs representing expressed mRNAs were isolated, sized, single-pass sequenced, and compared to known sequences in GenBank. Ninety-two clones provided sequence information for further analysis. Among these, 31 exhibited high homology to known genes. Three, 26S proteasomal ATPase (PSMC3), casein kinase 2 alpha subunit (CK2), and phosphoglycerate kinase (PGK) were selected and further characterized using real-time quantitative PCR to assess their differential expression in hatched blastocysts. Overall, a 1.3-, 1.6-, and 1.5-fold increase in expression level was observed in hatched blastocysts compared with intact blastocyst for PSMC3, CK2, and PGK, respectively. These results show that construction of subtracted cDNA libraries from small numbers of embryos is feasible and can provide information on gene expression patterns during preattachment embryogenesis.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0006-3363
pubmed:author
pubmed:issnType
Print
pubmed:volume
67
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
447-53
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:12135880-Adenosine Triphosphatases, pubmed-meshheading:12135880-Animals, pubmed-meshheading:12135880-Blastocyst, pubmed-meshheading:12135880-Casein Kinases, pubmed-meshheading:12135880-Cattle, pubmed-meshheading:12135880-Cloning, Molecular, pubmed-meshheading:12135880-DNA, Complementary, pubmed-meshheading:12135880-DNA Primers, pubmed-meshheading:12135880-Female, pubmed-meshheading:12135880-Fertilization in Vitro, pubmed-meshheading:12135880-Gene Expression Regulation, Developmental, pubmed-meshheading:12135880-Gene Library, pubmed-meshheading:12135880-Ovary, pubmed-meshheading:12135880-Phosphoglycerate Kinase, pubmed-meshheading:12135880-Pregnancy, pubmed-meshheading:12135880-Protein Kinases, pubmed-meshheading:12135880-RNA, pubmed-meshheading:12135880-RNA, Messenger, pubmed-meshheading:12135880-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:12135880-Sequence Homology, Nucleic Acid
pubmed:year
2002
pubmed:articleTitle
Analysis of gene expression in the bovine blastocyst produced in vitro using suppression-subtractive hybridization.
pubmed:affiliation
Department of Physiological Sciences, Division of Agricultural Science and Natural Resources, Oklahoma State University, Stillwater, OK 74078-2006, USA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't