Patent ID: 11866703
Assignee: LANZHOU VETERINARY RESEARCH INSTITUTE, CHINESE ACADEMY OF AGRICULTURAL SCIENCES
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 8:
9. The knockout method according to claim 1, wherein, after the electrotransformation, the method further comprises:
i. inoculating an obtained electrotransformation product into MDBK cells for cultivation, adding pyrimethamine at 6 h, and collecting host cells at 18 h;
ii. lysing and digesting the host cells to obtain cells with sporozoites;
iii. attacking chicks using the cells with sporozoites;
iv. collecting feces to obtain oocysts;
v. cultivating the oocysts into adult Eimeria tenella; 
vi. extracting genomic DNA (gDNA) of the adult Eimeria tenella; and
vii. with the gDNA as a template, conducting amplification using a PCR1 primer pair, a PCR2 primer pair, and a PCR3 primer pair, separately, wherein, when fragments can be obtained from both the amplification using the PCR1 primer pair and the amplification using the PCR2 primer pair, but no fragments can be obtained from the amplification using the PCR3 primer pair, the Eimeria tenella is NMT gene-knockout Eimeria tenella.