Title: Characterization of open reading frame-expressed sequence tags generated from Bos indicus and B-taurus mammary gland cDNA libraries

Author(s):  da Mota AF, Sonstegard TS, Van Tassell CP, Shade LL, Matukumalli LK, Wood DL, Capuco AV, Brito MAP, Connor EE, Martinez ML, Coutinho LL

Source:  ANIMAL GENETICS 35 (3): 213-219 JUN 2004

Document Type:  Article

Language:  English

Abstract:  Sequence-based gene expression data are used to interpret results from functional genomic and proteomics studies. Although more than 300 000 bovine-expressed sequence tags (ESTs) are available in public databases, a more thorough and directed sampling of the expressed genome is needed to identify new transcripts and improve assembly and annotation of existing transcript sequences. Accordingly, we examined the utility of constructing cDNA libraries synthesized by arbitrarily primed RT-PCR of mRNA from tissues not well represented in the publicly available bovine EST database. A total of 33 cDNA libraries were constructed from healthy and infected mammary gland tissues of Brazilian Gir and Holstein cattle. This series of libraries was used to generate 6481 open reading frame-expressed sequence tags (ORESTES) that assembled into 1798 unique sequence elements of which, 1157 did not significantly match sequence assemblies available in the Bos taurus gene index. However, a total of 264 of these 1157 sequence elements aligned with mouse and human expressed sequences demonstrating that ORESTES is an effective resource for discovery of novel expressed sequences in cattle. Furthermore, comparison of the alignment position of bovine ORESTES-derived sequence elements to human gene reference sequences suggested that the priming events for cDNA synthesis more often occurred at the central portion of a transcript, which may have contributed to the relatively high rate of novel sequence discovery.

Author Keywords:  bovine transcriptome; expressed sequence tags; mammary gland

KeyWords Plus:  HUMAN TRANSCRIPTOME; GENE INDEX

Addresses:  Sonstegard TS (reprint author), ARS, USDA, Bovine Funct Genom Lab, Beltsville Agr Res Inst BARC E, Beltsville, MD 20705 USA
ARS, USDA, Bovine Funct Genom Lab, Beltsville Agr Res Inst BARC E, Beltsville, MD 20705 USA
EMBRAPA, Gado Leite, Natl Dairy Cattle Res Ctr, Bairro Dom Bosco, Juiz de Fora, MG Brazil
Univ Sao Paulo, ESALQ, Dept Zootecnia, Lab Anim Biotechnol, Piracicaba, SP Brazil
George Mason Univ, SCS, Manassas, VA USA

E-mail Addresses: tads@anri.barc.usda.gov

Publisher:  BLACKWELL PUBLISHING LTD, 9600 GARSINGTON RD, OXFORD OX4 2DG, OXON, ENGLAND

Subject Category:  AGRICULTURE, DAIRY & ANIMAL SCIENCE; GENETICS & HEREDITY

IDS Number:  821UL

ISSN:  0268-9146