Molecular cloning and expression of the CRISP family of proteins in the boar

Biology of Reproduction
Melissa L VadnaisK P Roberts

Abstract

The family of mammalian cysteine-rich secretory proteins (CRISP) have been well characterized in the rat, mouse, and human. Here we report the molecular cloning and expression analysis of CRISP1, CRISP2, and CRISP3 in the boar. A partial sequence published in the National Center for Biotechnology Information (NCBI) database was used to derive the full-length sequences for CRISP1 and CRISP2 using rapid amplification of cDNA ends. RT-PCR confirmed the expression of these mRNAs in the boar reproductive tract, and real time RT-PCR showed CRISP1 to be highly expressed throughout the epididymis, with CRISP2 highly expressed in the testis. A search of the porcine genomic sequence in the NCBI database identified a BAC (CH242-199E6) encoding the CRISP1 gene. This BAC is derived from porcine Chromosome 7 and is syntenic with the regions of the mouse, rat, and human genomes encoding the CRISP gene family. This BAC was found to encode a third CRISP protein with a predicted amino acid sequence of high similarity to human CRISP3. Using RT-PCR we show that CRISP3 expression in the boar reproductive tract is confined to the prostate. Recombinant porcine (rp) CRISP2 protein was produced and purified. When incubated with capacitated boar sperm, ...Continue Reading

References

Jan 1, 1992·Mammalian Genome : Official Journal of the International Mammalian Genome Society·N MizukiM Kasahara
Nov 1, 1980·Biology of Reproduction·A C KohaneJ A Blaquier
Feb 28, 1997·Journal of Molecular Biology·C FernándezK Wüthrich
Jan 15, 1998·European Journal of Biochemistry·B HaendlerW D Schleuning
Jun 11, 1998·Molecular Reproduction and Development·M K O'BryanD M de Kretser
Sep 28, 1998·Biochimica Et Biophysica Acta·A SchambonyE Töpfer-Petersen
Jan 20, 1999·Proceedings of the National Academy of Sciences of the United States of America·R L BrownJ W Crabb
Feb 16, 1999·Journal of Cellular Physiology·B HaendlerW D Schleuning
Jan 6, 2001·Molecular Reproduction and Development·M K O'BryanD M De Kretser
Jun 6, 2002·European Journal of Biochemistry·Yasuo YamazakiTakashi Morita
Jul 29, 2006·Developmental Biology·Diego A EllermanPatricia S Cuasnicú
Oct 31, 2006·Biochemical and Biophysical Research Communications·Feng WangTao Jiang
Jan 5, 2007·Biology of Reproduction·Dolores BussoPatricia S Cuasnicú
May 1, 2007·Biochemical and Biophysical Research Communications·Naoko ItoShoji Odani
Sep 12, 2007·Bioinformatics·M A LarkinD G Higgins

❮ Previous
Next ❯

Citations

Sep 28, 2015·Toxicon : Official Journal of the International Society on Toxinology·Félix A UrraAntonieta Labra
Jun 15, 2012·Reproduction in Domestic Animals = Zuchthygiene·I CaballeroJ M Vázquez
Jun 10, 2020·BMC Evolutionary Biology·Lena ArévaloEduardo R S Roldan
Mar 7, 2020·Cell and Tissue Research·Michal ZigoPeter Sutovsky
May 29, 2021·Journal of Trace Elements in Medicine and Biology : Organ of the Society for Minerals and Trace Elements (GMS)·Parisa DolatiIman Jamhiri
Jun 22, 2021·Theriogenology·Manita WittayaratSaritvich Panyaboriban

❮ Previous
Next ❯

Methods Mentioned

BETA
protein assay
PCR
Assay
transfection
electrophoresis
Genotyping
flow cytometry

Software Mentioned

CLUSTAL W
CLUSTALW
FlowJo
Statistical Analysis Systems
Applied 7500 Fast SDS
BLAST

Related Concepts

Related Feeds

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.