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Auxin-induced mRNA species in tobacco cell cultures 总被引:6,自引:0,他引:6
E. J. van der Zaal J. Memelink A. M. Mennes A. Quint K. R. Libbenga 《Plant molecular biology》1987,10(2):145-157
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D. -D. Sui J. -L. Wu H. Zhang H. Li Z. -M. Zhou D. -H. Zhang C. -X. Han 《Molecular Biology》2014,48(5):646-654
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. 相似文献