We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Molecular cloning, structural analysis, and expression of zona pellucida glycoprotein ZP3 gene from Chinese zokor, Myospalax fontanierii.
- Authors
Sui, D.; Wu, J.; Zhang, H.; Li, H.; Zhou, Z.; Zhang, D.; Han, C.
- Abstract
The zona pellucida 3 (ZP3) plays a crucial role in reproductive immunology. We obtained a full-length cDNA encoding Chinese zokor ZP3, using rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR). The cDNA contains an open reading frame of 1269 nucleotides encoding a polypeptide of 422 amino acid residues. The amino acid sequence has a high degree of homology with those of hamster (78%), mouse (76%), and rat (74%). XhoI and SacI sites after restriction give an1158 bp fragment of zokor ZP3 cDNA, excluding the signal sequence and transmembrane-like domain, which was cloned under the phage T7 promoter-lac operator control in the pET-28a(+) vector. Recombinant pET-zokorZP3(r-ZP3) was expressed as a poly-histidine fusion protein in E. coli strain BL21 (DE3). Optimum expression of r-ZP3 was observed at 28°C, 1 mM IPTG and 2 h of inducing. The purified protein was tested by Western blot.
- Subjects
ZONA pellucida; ZOKORS; LABORATORY rodents; MOLECULAR cloning; GENE expression in mammals; ANTISENSE DNA
- Publication
Molecular Biology, 2014, Vol 48, Issue 5, p646
- ISSN
0026-8933
- Publication type
Article
- DOI
10.1134/S0026893314050148