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Generation of Multi-Transgenic Pigs Using PiggyBac Transposons Co-expressing Pectinase, Xylanase, Cellulase, β-1.3-1.4-Glucanase and Phytase

The current challenges facing the pork industry are to maximize feed efficiency and minimize fecal emissions. Unlike ruminants, pigs lack several digestive enzymes such as pectinase, xylanase, cellulase, β-1.3-1.4-glucanase, and phytase which are essential to hydrolyze the cell walls of grains to re...

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Autores principales: Wang, Haoqiang, Li, Guoling, Zhong, Cuili, Mo, Jianxin, Sun, Yue, Shi, Junsong, Zhou, Rong, Li, Zicong, Wu, Zhenfang, Liu, Dewu, Zhang, Xianwei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7734351/
https://www.ncbi.nlm.nih.gov/pubmed/33329743
http://dx.doi.org/10.3389/fgene.2020.597841
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author Wang, Haoqiang
Li, Guoling
Zhong, Cuili
Mo, Jianxin
Sun, Yue
Shi, Junsong
Zhou, Rong
Li, Zicong
Wu, Zhenfang
Liu, Dewu
Zhang, Xianwei
author_facet Wang, Haoqiang
Li, Guoling
Zhong, Cuili
Mo, Jianxin
Sun, Yue
Shi, Junsong
Zhou, Rong
Li, Zicong
Wu, Zhenfang
Liu, Dewu
Zhang, Xianwei
author_sort Wang, Haoqiang
collection PubMed
description The current challenges facing the pork industry are to maximize feed efficiency and minimize fecal emissions. Unlike ruminants, pigs lack several digestive enzymes such as pectinase, xylanase, cellulase, β-1.3-1.4-glucanase, and phytase which are essential to hydrolyze the cell walls of grains to release endocellular nutrients into their digestive tracts. Herein, we synthesized multiple cellulase and pectinase genes derived from lower organisms and then codon-optimized these genes to be expressed in pigs. These genes were then cloned into our previously optimized XynB (xylanase)- EsAPPA (phytase) bicistronic construct. We then successfully generated transgenic pigs that expressed the four enzymes [Pg7fn (pectinase), XynB (xylanase), EsAPPA (phytase), and TeEGI (cellulase and β-glucanase)] using somatic cell cloning. The expression of these genes was parotid gland specific. Enzymatic assays using the saliva of these founders demonstrated high levels of phytase (2.0∼3.4 U/mL) and xylanase (0.25∼0.42 U/mL) activities, but low levels of pectinase (0.06∼0.08 U/mL) activity. These multi-transgenic pigs are expected to contribute to enhance feed utilization and reduce environmental impact.
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spelling pubmed-77343512020-12-15 Generation of Multi-Transgenic Pigs Using PiggyBac Transposons Co-expressing Pectinase, Xylanase, Cellulase, β-1.3-1.4-Glucanase and Phytase Wang, Haoqiang Li, Guoling Zhong, Cuili Mo, Jianxin Sun, Yue Shi, Junsong Zhou, Rong Li, Zicong Wu, Zhenfang Liu, Dewu Zhang, Xianwei Front Genet Genetics The current challenges facing the pork industry are to maximize feed efficiency and minimize fecal emissions. Unlike ruminants, pigs lack several digestive enzymes such as pectinase, xylanase, cellulase, β-1.3-1.4-glucanase, and phytase which are essential to hydrolyze the cell walls of grains to release endocellular nutrients into their digestive tracts. Herein, we synthesized multiple cellulase and pectinase genes derived from lower organisms and then codon-optimized these genes to be expressed in pigs. These genes were then cloned into our previously optimized XynB (xylanase)- EsAPPA (phytase) bicistronic construct. We then successfully generated transgenic pigs that expressed the four enzymes [Pg7fn (pectinase), XynB (xylanase), EsAPPA (phytase), and TeEGI (cellulase and β-glucanase)] using somatic cell cloning. The expression of these genes was parotid gland specific. Enzymatic assays using the saliva of these founders demonstrated high levels of phytase (2.0∼3.4 U/mL) and xylanase (0.25∼0.42 U/mL) activities, but low levels of pectinase (0.06∼0.08 U/mL) activity. These multi-transgenic pigs are expected to contribute to enhance feed utilization and reduce environmental impact. Frontiers Media S.A. 2020-11-30 /pmc/articles/PMC7734351/ /pubmed/33329743 http://dx.doi.org/10.3389/fgene.2020.597841 Text en Copyright © 2020 Wang, Li, Zhong, Mo, Sun, Shi, Zhou, Li, Wu, Liu and Zhang. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Genetics
Wang, Haoqiang
Li, Guoling
Zhong, Cuili
Mo, Jianxin
Sun, Yue
Shi, Junsong
Zhou, Rong
Li, Zicong
Wu, Zhenfang
Liu, Dewu
Zhang, Xianwei
Generation of Multi-Transgenic Pigs Using PiggyBac Transposons Co-expressing Pectinase, Xylanase, Cellulase, β-1.3-1.4-Glucanase and Phytase
title Generation of Multi-Transgenic Pigs Using PiggyBac Transposons Co-expressing Pectinase, Xylanase, Cellulase, β-1.3-1.4-Glucanase and Phytase
title_full Generation of Multi-Transgenic Pigs Using PiggyBac Transposons Co-expressing Pectinase, Xylanase, Cellulase, β-1.3-1.4-Glucanase and Phytase
title_fullStr Generation of Multi-Transgenic Pigs Using PiggyBac Transposons Co-expressing Pectinase, Xylanase, Cellulase, β-1.3-1.4-Glucanase and Phytase
title_full_unstemmed Generation of Multi-Transgenic Pigs Using PiggyBac Transposons Co-expressing Pectinase, Xylanase, Cellulase, β-1.3-1.4-Glucanase and Phytase
title_short Generation of Multi-Transgenic Pigs Using PiggyBac Transposons Co-expressing Pectinase, Xylanase, Cellulase, β-1.3-1.4-Glucanase and Phytase
title_sort generation of multi-transgenic pigs using piggybac transposons co-expressing pectinase, xylanase, cellulase, β-1.3-1.4-glucanase and phytase
topic Genetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7734351/
https://www.ncbi.nlm.nih.gov/pubmed/33329743
http://dx.doi.org/10.3389/fgene.2020.597841
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