Category Archives: Ovarian Follicle

Cloning, Sequencing, and Localization of Bovine: DISCUSSION(3)

Therefore, ERp may play an important role in ovarian follicular growth, development, and subsequent ovulation. Immunohistochemistry for ERp revealed that there may be a slight amount of ERp protein expression within the theca cells of bovine antral follicles, whereas on … Continue reading

Cloning, Sequencing, and Localization of Bovine: DISCUSSION(2)

Finally, the initially reported rat ERp start codon corresponds to bERp amino acid #46, a valine, and thus it cannot be the initiation codon for bERp (Fig. 2, a and b). buy yasmin online Overall, bERp protein is most homologous … Continue reading

Cloning, Sequencing, and Localization of Bovine: DISCUSSION(1)

In these experiments, the complete protein coding sequence of bERp mRNA was determined. In situ hybridization and immunohistochemistry localized bERp mRNA and protein to the granulosa cells of small, medium, and large antral follicles. buy ventolin inhalers Bovine and human … Continue reading

Cloning, Sequencing, and Localization of Bovine: RESULTS(2)

Bovine ERp Protein Localization within Ovarian Follicles Immunohistochemical localization of ERp within the bovine ovary detected ERp protein within the granulosa cells of small (Fig. 4A), medium (Fig. 4C), and large (Fig. 4E) antral follicles. Positive staining for ERp was … Continue reading

Cloning, Sequencing, and Localization of Bovine: RESULTS(1)

Bovine ERp Sequence As evidenced by sequence analysis, bERp mRNA spanned an open reading frame of 1584 nucleotides encoding a protein of 527 amino acids (Fig. 1). Comparison of the derived bERp amino acid sequence with rat, GenBank accession no. … Continue reading

Cloning, Sequencing, and Localization of Bovine: MATERIALS AND METHODS(5)

Immunohistochemistry Immunohistochemistry for ERp was performed as previously described. Briefly, paraffin-embedded bovine ovarian follicle sections (4-5 ^m) were placed in a microwave for 5-10 min to expose the cellular antigens and increase binding affinity of the antibody to the antigen. … Continue reading

Cloning, Sequencing, and Localization of Bovine: MATERIALS AND METHODS(4)

Restriction digest and subsequent in vitro transcription, using purified linearized cDNA templates, were performed to generate both antisense and sense probes. BsgI restriction enzyme and T3 polymerase were used to generate the first antisense bERp probe, which spanned exons 2-5, … Continue reading

Cloning, Sequencing, and Localization of Bovine: MATERIALS AND METHODS(3)

DNA Sequencing and Analysis RT-PCR-amplified bERp cDNA fragments were cloned into the pCR-Script AMP (SK[+]) (Stratagene, La Jolla, CA) vector system. Gel purified-cDNA and plasmid DNA were sequenced using the cycle sequencing fluorometric method (PE Applied Biosystems, Foster City CA). … Continue reading

Cloning, Sequencing, and Localization of Bovine: MATERIALS AND METHODS(2)

Each reaction tube contained a final concentration of 0.2 mM dNTPs (Promega, Madison, WI), 5 mM dithiothreitol (Boehringer-Mannheim), 1.6 U RNasin (Promega), single-strength RT-PCR buffer with 1.5 mM Mg2+ mix (Boehringer-Mannheim), and enzyme mix of avian myeloblastosis virus reverse transcriptase … Continue reading

Cloning, Sequencing, and Localization of Bovine: MATERIALS AND METHODS(1)

Tissue Collection Holstein dairy cows used for these experiments were initially housed on a dry feedlot, and from the onset of estrus they were housed inside the Animal Science Research Center large-animal facility. Ovarian activity was monitored by transrectal ultrasound … Continue reading