Production of cleavage-resistant phytase transgenic pigs by handmade cloning

文献类型： 外文期刊

第一作者： Zhang, Mingyuan

作者： Zhang, Mingyuan;Zhang, Mingyuan;Yang, Haowen;Chen, Shijin;Chen, Xiaoliang;Huang, Yuemeng;Chen, Baojian;Wu, Yanjun;Jiang, Qinyang;Guo, Yafen;Wei, Yingming;Lan, Ganqiu;Liang, Jing;Jiang, Hesheng;Zhang, Mingyuan;Zhang, Jun;Guo, Xiaoping;Yang, Haowen;Chen, Xiaoliang;Lin, Xiukun

作者机构：

关键词： Cleavage-resistant phytase; Handmade cloning; Transgenic pigs

期刊名称：THERIOGENOLOGY （ 影响因子：2.8； 五年影响因子：2.6 ）

ISSN： 0093-691X

年卷期： 2024 年 224 卷

页码：

收录情况： SCI

摘要： With the rapid development of intensive animal husbandry in the livestock industry, large quantities of manure waste containing phytate phosphorus are being generated. Phytase can effectively solve the problem of high phosphorus pollution in the feces of monogastric animals. Enviropig, which produces phytase in the salivary glands and secretes the enzyme in the saliva, were first generated in 1999. However, phytase is easily inactivated during digestion. To address this problem, cleavage-resistant phytase transgenic pigs were generated using handmade cloning in this study. Transgene construction was improved and three cell lines carrying Cafp were obtained. In total, 810 blastocysts were generated and 712 good-quality were transferred into six recipients. Fourteen piglets were born, of which six survived after weaning. Polymerase chain reaction and sequencing results showed that seven (three live and four dead) of the fourteen piglets carried Cafp . Phytase activity in the saliva of the six live cloned pigs was tested at four months of age, and only one pig had 0.155 FTU/mL enzyme activity. The other five pigs may not have been activated in the transgenic parotid gland. Among all the transgenic pigs, the highest phosphorus digestion rate was 59.2% of intake, representing a 25.4% decrease in fecal emission compared to the average of controls. Immunohistochemical results on the three Cafp- positive pigs that died after six months of age showed that the transgene was only expressed in parotid glands, confirming tissue-specific gene expression. In conclusion, cleavage-resistant phytase transgenic pigs were successfully produced through handmade cloning. The cloned pigs offer a unique biological approach to managing phosphorus nutrition and environmental pollution in animal husbandry.

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