Wyniki wyszukiwania - Biblioteka Nauki

Ten serwis zostanie wyłączony 2025-02-11.

Nowa wersja platformy, zawierająca wyłącznie zasoby pełnotekstowe, jest już dostępna.
Przejdź na https://bibliotekanauki.pl

Ograniczanie wyników

3 Animal Science Papers and Reports

Znaleziono wyników: 3

Liczba wyników na stronie

Wyniki wyszukiwania

Sortuj według:

Ogranicz wyniki do:

Selection of reference genes for gene expression studies in porcine hepatic tissue using quantitative real-time polymerase chain reaction

100%

Pierzchala M. , Pareek C. S. , Urbanski P. , Goluch D. , Kamyczek M. , Rozycki M. , Kuryl J.

Animal Science Papers and Reports

2011

tom 29

nr 1

Quantitative real-time polymerase chain reaction (RT-qPCR) has become an indispensable technique for accurate determination of gene expression in variety of samples. Accurate and reliable quantification, however, depends on a proper normalization strategy. Normalization with multiple uniformly expressed reference genes is becoming the standard, although the most suitable reference genes dependent on the used experimental factors as well as the tissue or cell type studied. In this study, the stability of various reference genes was investigated in porcine hepatic tissue. The study was conducted on Polish Large White, Polish Landrace, Pietrain, Pulawska and Duroc pigs slaghtered at different ages. Nine reference genes (ACTB, B2M, GAPDH, HPRT1, RPL13A, SDHA, TBP, TOP2B and YWHAZ) were investigated on 180 mRNA samples of porcine hepatic tissue. Based on geNorm and NormFinder analysis, three most stable (HPRT1, TOP2B and TBP) and three moderately (GAPDH, ACTB and SDHA) stable reference genes were identified. The study provides a new panel of reference genes for normalization of the expression of a gene of interest in porcine liver tissue. It is concluded that the use of a single gene for normalization may lead to relatively large errors, so it is important to use multiple control genes based on a survey of potential reference genes applied to gene expression profiling studies of candidate genes for economic traits in pigs.

Expression profile of MYF5 and MYF6 genes in skeletal muscles of young growing gilts of five breeds at different ages, based on the most stable reference genes

84%

Pierzchala M. , Pareek C. S. , Lisowski P. , Urbanski P. , Goluch D. , Czarnik U. , Kamyczek M. , Rozycki M. , Cooper R. G. , Kuryl J.

Animal Science Papers and Reports

2011

tom 29

nr 3

The expression profile was evaluated of MYF5 and MYF6 genes in skeletal muscles of young growing Polish Large White (PLW), Polish Landrace (PL), Pietrain (PIE), Duroc (DUR) and Pulawska (PUL) gilts at different ages. Normalization of MYF5 and MYF6 was performed on reliable porcine reference genes (PRGs), where expression stabilities of nine of them (ACTB, B2M, GAPDH, SDHA, HPRTI, RPL13A, YWHAZ, TBP, TOP2B) were evaluated by RT-qPCR method and NormFinder software. Results revealed HPRTI, TBP and TOP2B as highly stable and PRGs. The age-dependent and breed-specific skeletal muscle expression comparisons revealed highly significant (P<0.01) differences in MYF6 expression levels of all skeletal muscles among investigated breeds. MYF6 gene expression in PIE and DUR were higher compared to PLW, PL and PUL gilts. Contrarily, paired-wise comparison of MYF5 gene expression showed only significant difference between DUR and PUL for semimembranosus, and PL and PLW, DUR and PL, PIE and PL, DUR and PUL and PIE and PUL for gluteus medius muscle. There was no significant relationship identified between gilt ages and the level of expression of MYF5 and MYF6 genes. However, their highest expression was identified in longissimus dorsi followed by gluteus medius and semimembranosus muscles. It is concluded that normalization of gene expression has to be done on more than one PRG to reduce the errors in transcription level estimates. Moreover, significantly different breed-specific expression of porcine MYF5 and MYF6 allowed the authors to prioritize these genes as potential candidate genes for trait-associated study.

Gene expression profiling by cDNA-AFLP and identification of differentially expressed transcripts of bovine pituitary gland in growing bulls of dairy breeds

67%

Pareek C. S. , Pierzchala M. , Michno J. , Szymanska S. , Smoczynski R. , Jasik N. , Golebiewski M. , Tyburski J. , Urbanski P. , Goluch D. , Oprzadek J. , Zwierzchowski L. , Siriluck P. , Wimmers K.

tom 30

nr 2

The study utilized age-dependent breed-specific gene expression profiling by cDNA-AFLP technique in identification of differentially expressed (DE) candidate genes for growth and development trait. Results revealed 15579 transcript derived fragments (TDFs) expressed in bovine pituitary gland tissue using 96 unique PCR reactions with primer combinations (PC) of TaqI-MseI for Polish Holstein (PH) and Polish Red (PR) dairy breeds. Gene expression profiling by cDNA-AFLP identified 1451,2877 and 4084 identically displayed (iDD), differentially displayed (DD) and single displayed (sDD) TDFs, respectively. In all transcript profiles, frequencies of DD TDFs were higher than that of iDD TDFs. A total of 60 DD TDFs bands were excised for PH (n=40) and PR (n=20) . Direct sequencing results revealed identification of 24 and 8 DE-TDFs sequences in Polish HF and PR. Based on the significance of BLAST and sequence alignment score, analysis 12 and 4 DE-TDFs sequences of PH and PR were excluded. Overall, 12 Polish HF and 4 PR breed specific DE-TDFs sequence expressed in bovine pituitary gland tissue were identified. The identified DE-TDFs sequences were represented in all bovine chromosomes, except BTA3, 4, 6, 7, 8, 9, 11, 12, 17, 18, 21, 22, 23, 25 and 26. TDF annotation results identified eight sequences that have BLAST hits to known annotated bovine genes and eight sequences to unannotated contig regions in latest gene ensemble database Btau 4,0. Two breed-specific target genes i.e., bovine glycophorin C (PH) and bovine arachidonate 5-lipoxygenase (Alox5) isoform 1 (PR) were validated by qRT-PCR. Within breed the age-dependent qRT-PCR analysis revealed that expression levels were differed significantly high (P<0.0001) with nine folds higher expression in young bulls at the age of 6 month (glycophorin C) and 9 month (Alox5 isoform 1). Between breed the qRT-PCR analyses revealed that the expression levels were highly significant for glycophorin C gene in PR and Alox5 isoform 1 gene in both breeds. It was concluded that gene expression profiling by cDNA-AFLP is a reliable technique for identification of trait-associated DE candidate genes, and helpful in elucidating and understanding molecular basis of postnatal growth and development of cattle.