SOD2 Mouse Model Building And Identification

Objective: Familiar with the Talens one of three categories genome editing nuclease,construction of SOD2 transgenic mouse model by using Tanlens system,the homozygous model of SOD2 transgenic mice was obtained by breeding system and later identification.Methods:(1)With the wild type C57 mice as experimental subjects,we constructed SOD2 transgenic mouse model in Talens system,and verified by conventional PCR and 1% agarose gel electrophoresis system.(2)The construction of the heterozygous SOD2 transgenic mice and wild-type C57 mice were bred by conventional PCR,and 1% agarose gel electrophoresis system screened positive expression in mice,and the use of positive mouse breeding lines to the wild type C57 mice.(3)When the strain reaches a certain size,each group choose positive results of two mice and wild-type C57 mice were screened by Wb and Q-PCR two is more suitable for the backcross lines,then backcross of SOD2 transgenic mice until compatriots homozygous.Results:(1)Through the conventional PCR and 1% agarose gel electrophoresis system,we successfully obtained heterozygous SOD2 transgenic black mice of 10.(2)Finally,6 strains,No.2,No.4,No.7,No.9,which had the requirements of backcross,were successfully obtained by breeding,and there were only about 1strains in each of them,including 8,10,9,10,8,and 9 positive mice.(3)The number of positive mice 6 lines are about 10,6 lines are in conformity with the requirements of the SOD2 backcross mice,Q-PCR results,2 strains of 6 strains and 7strains of the most suitable fellow compatriots backcross backcross,final 2 strains,7were homozygous the SOD2 transgenic mice.Conclusion:(1)Talens system can complete the construction of SOD2 transgenic mice.(2)through the normal breeding lines,coupled with the conventional PCR.1% agarose gel electrophoresis,Western blotting,Q-PCR technology can complete the identification of transgenic mice with SOD2 genotype,and transgenic mice for SOD2 to get laid backcross homozygotes based on the technical level.