Cloning and expression pattern of erk2 gene in Inner Mongolia Cashmere goat.
- Author:
Yanfeng WANG
;
Manlin WU
;
Xiaojing WANG
;
Jing WANG
;
Yang LI
;
Mengyao LIAN
;
Zhigang WANG
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Animals;
China;
Cloning, Molecular;
DNA, Complementary;
genetics;
Goats;
genetics;
Male;
Mitogen-Activated Protein Kinase 1;
genetics;
metabolism;
Molecular Sequence Data;
Real-Time Polymerase Chain Reaction;
Testis;
metabolism
- From:
Chinese Journal of Biotechnology
2013;29(12):1743-1752
- CountryChina
- Language:Chinese
-
Abstract:
The study aims at cloning the CDS fragment of erk2 gene cDNA in Inner Mongolia Cashmere Goat and analyzing its tissue-specific expression, erk2 gene cDNA was cloned by RT-PCR. The nucleotide sequence was analyzed by Blast and amino acid sequence was analyzed by online softwares SMART and Psite. The tissue-specific expression pattern of erk2 was analyzed by quantitative RT-PCR. The expression of erk2 in testis of goat was detected by Immunohistochemistry. The cloned erk2 gene cDNA (GenBank Accession No. JX569765) was 1 083 bp in length, including a complete ORF encoding 360 amino acids residues. The amino acid sequence shares 100% identity with the Bos Taurus ERK2 (Bos Taurus BC133588.1). Analysis by SMART suggests that the encoded protein contained a "TEY" structure and an S-TKc domain possessing serine/threonine kinase catalytic activity. Analysis with Psite indicates one cAMP-/cGMP-dependent protein kinase phosphorylation site, 3 protein kinase C phosphorylation sites, 5 casein kinase II phosphorylation sites, 2 protein kinases ATP-binding region signatures and one serine/threonine protein kinases active-site signature in this protein. Analysis by Psort (k-NN prediction) suggestes that this protein most probably is localized in cytoplasm. The results of quantitative RT-PCR show that the expression of erk2 mRNA was higher in heart, skin and breast, whereas lower in spleen and kidney. ERK2 protein was detected in testis by immunohistochemistry.
