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Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells

Journal of Integrative Agriculture 2015年第4期14卷 732-740页

作者： ZHANG Xian-wei LI Zi-cong MENG Fan-ming WANG De-hua LIU De-wu HE Xiao-yan SUN Yue BAI Yin-shan WU Zhen-fang National Engineering Research Center for Breeding Swine Industry South China Agricultural University Wen's Research Institute Guangdong Wen's Food Group Co.Ltd.

Many animal feed grains contain high β-glucan in the cell wall. Pigs do not secret β-glucanase to degrade the β-glucan in their feed. The indigestible β-glucan not only blocks the release of nutrients from the grain cell wall, but also increases the digesta viscosity in the gastrointestinal tract of pigs. Therefore, dietary β-glucan significantly inhibits nutrient digestion and absorption in pigs. Transgenic expression of β-glucanase in the digestive tract of pigs may offer a solution to solve this problem. In the current study, four artificial codon-optimized β-glucanases genes was prepared and expressed in porcine cells. Only p Bg A and p Egx showed high activity in transfected pig kidney cells. To improve the p H range and p H stability of β-glucanase, the two β-glucanases, p Bg A and p Egx, were co-expressed in pig kidney cells and salivary gland cells by Linker A3 or 2A peptide. The resulting dual enzymes of p Bg A3 p Eg and p Bg2 Ap Eg showed significantly enlarged p H range and significantly increased p H stability, as compared to parental enzymes. These results provide useful data for future study on increasing the feed digestibility of pigs by transgenic expression of β-glucanase in their salivary glands.

关键词： β-glucanase bicistronic pig feed digestibility salivary gland cells transgenic

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IRES-mediated cap-independent translation,a path leading to hidden proteome

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Journal of Molecular Cell Biology 2019年第10期11卷 911-919页

作者： Yun Yang Zefeng Wang CAS Key Laboratory of Computational Biology Biomedical Big Data CenterCAS-MPG Partner Institute for Computational BiologyShanghai Institute of Nutrition and HealthChinese Academy of SciencesShanghai 200031China University of Chinese Academy of Sciences Chinese Academy of SciencesShanghai 200031China CAS Center for Excellence in Molecular Cell Science Chinese Academy of SciencesShanghai 200031China

Most eukaryotic mRNAs are translated in a cap-dependent fashion;however,under stress conditions,the cap-independent translation driven by internal ribosomal entry sites(IRESs)can serve as an alternative mechanism for protein *** IRESs have been discovered from viral or cellular mRNAs to promote ribosome assembly and initiate translation by recruiting different trons-acting *** the mechanisms of translation initiation driven by viral IRESs are relatively well understood,the existence of cellular IRESs is still under debate due to the limitations of translation reporter systems used to assay IRES activities.A recent screen identified>1000 putative IRESs from viral and human mRNAs,expanding the scope and mechanism for capindependent ***,a large number of circular RNAs lacking free ends were identified in eukaryotic cells,many of which are found to be translated through *** findings suggest that IRESs may play a previously unappreciated role in driving translation of the new type of mRNA,implying a hidden proteome produced from cap-independent translation.

关键词： IRES ITAF translation bicistronic system circular RNA

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