Establishing Prokaryotic Expression System of Angiotensin-Converting Enzyme 2 (ACE2) gene in pigs

Hang Xiao; Xin-Tian Nie; Xiao-Xia Ji; Shu-ping Yan; Bin Zhu; Yuan-Shu Zhang

So far, the research on ACE2 has been focused on rodents and human beings. 53 The research gaps in other animals, especially production animals, such as cattle, 54 sheep, pigs still need to be filled. Particularly, GenBank still lacks the information of The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.12.988634 doi: bioRxiv preprint 61 polyclonal antibody was obtained and the expression distribution of ACE2 was 62 determined. We hope to be able to provide the whole sequence of ACE2 gene and other 63 related information for Genbank. 75 The Healthy newborn piglets were terminally anesthetized via an intraperitoneal 76 (ip.) injection of 20% urethane (5 mL/kg body weight), were decapitated within 15 77 min after anesthesia took effect (i.e. animals lost conciseness and whole body muscle 78 relaxation was observed), and all tissues including heart, liver, hung, kindey, stomach 79 and soon were collected and frozen immediately in liquid nitrogen and then stored at 80 -80°C , Part of tissues were fixed immediately with 4% buffered paraformaldehyde 81 for 24 hours, then transferred to 20% sucrose solution for 3 h. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.12.988634 doi: bioRxiv preprint Freund's adjuvant. Serum was collected from heart method and stored at −80 o C. 152 The protein of heart, spleen, kidney, liver, duodenum, liver of pig, and also liver 153 and kidney tissue of chicken were extracted from Roe. The protein concentration of 154 each tissue was measured by BCA kit(Beyotime Biotechnology, China). The IPEC-J2 cells were cultured in 6 orifice plates with cell crawling slices.