Abstract:
The primers were designed and synthesized based on the nucleotide sequences of porcine Th1/Th2 cytokinesavailable in GenBank .The 101 bp fragments were amplified by RT -PCR from the porcine peripheral bloodmononuclear cells(PBMC), and then the fragments were cloned and sequenced .The recombinant plasmids were diluted by 10 -fold serial and used as the real -time PCR standard templates .Their standard curves and the corresponding linearregression equations were obtained .The results indicated that the standard curves were shown to be of high linearity ,specificity,sensitivity and reproducibility,and they provided powerful tools for quantification of IL-2,IL -12,IFN -γ,IL-4,IL -6 and IL-10 mRNA in pigs.
Key words:
Thl/Th2cytokines,
PBMC,
Recombinantplasmi
CLC Number:
WENLi-bin, HEKong-wang, YANGHan-chun, GUORong-li, LICheng-ren, LUWei-cai, JIAFu-cong. Establishment of Standard Curves for Detection of PorcineThl/Th2 Cytokines Genes Recombinant Plasmids UsingReal-timePCR[J]. ACTA AGRICULTURAE BOREALI-SINICA, doi: 10.7668/hbnxb.2009.S1.002.