Method for screening feed antibiotic substitutes and products thereof
A technology of antibiotics and substitutes, applied in biochemical equipment and methods, material inspection products, chemical instruments and methods, etc., can solve the problems of long screening and identification cycle, low efficiency, high cost, etc.
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Embodiment 2
[0086] Example 2. Assessment of the Antibiotic Potential of Alpha-Ketoglutarate
[0087] The stably transformed PXR porcine intestinal epithelial cell line pxr_IPEC-J2 obtained by the method described in the patent was used as the research object to evaluate the substitution potential of alpha-ketoglutarate. The specific implementation is as follows: 1) The stable transfection cell line pxr_IPEC-J2 is cultured in the whole medium, and the cells are cultured to the logarithmic phase (80-90% fusion) for subculture and plating. 2) After the cells grow to the logarithmic phase (80-90% confluence), the experimental group is treated with alpha-ketoglutarate at different concentrations (1.0 mM, 2.0 mM, 5.0 mM) for 48 hours. 3) Collecting cells, extracting mRNA and total cell protein respectively. 4) Real time PCR technology was used to detect the expression of pgPXRmRNA in porcine intestinal cell lines. Second, the expression of pgPXR protein was detected by western blot technique....
Embodiment 3
[0088] Example 3. Assessment of the Antibiotic Potential of Alliin (Precursor of Allicin)
[0089] The stably transformed PXR porcine intestinal epithelial cell line pxr_IPEC-J2 obtained by the method described in the patent was used as the research object to evaluate the alliinin substitution potential. The specific implementation is as follows: 1) The stable transfection cell line pxr_IPEC-J2 is cultured in the whole medium, and the cells are cultured to the logarithmic phase (80-90% fusion) for subculture and plating. 2) When the cells grow to the logarithmic phase (80-90% fusion), the experimental group is added with different concentration gradients (1.0ug / mL, 2.0ug / mL, 5.0ug / mL,) of alliin, and the control group is added with the experimental The same amount of DMSO was treated for 48 hours. 3) Collecting cells, extracting mRNA and total cell protein respectively. 4) Real time PCR technology was used to detect the expression of pgPXR mRNA in porcine intestinal cell lin...
Embodiment 4
[0090] Embodiment 4, the potential evaluation of the replacement potential of Boluohui
[0091] The stably transformed PXR porcine intestinal epithelial cell line pxr_IPEC-J2 obtained by the method described in the patent was used as the research object to evaluate the potential of replacing Boluohui. The specific implementation is as follows: 1) The stable transfection cell line pxr_IPEC-J2 is cultured in the whole medium, and the cells are cultured to the logarithmic phase (80-90% fusion) for subculture and plating. 2) When the cells grow to the logarithmic phase (80-90% confluence), the experimental group is added with different concentration gradients (1.0ug / mL, 2.0ug / mL, 5.0ug / mL,) and the control group is added to the experimental group. The same amount of DMSO was treated for 48 hours. 3) Collecting cells, extracting mRNA and total cell protein respectively. 4) Real time PCR technology was used to detect the expression of pgPXR mRNA in porcine intestinal cell lines. ...
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