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AIM:To clone and express mouse peroxiredoxin I in IEC-6cells.METHODS:Total RNAs were isolated from cultured IEC-6cells,and the coding region of peroxiredoxin I was amplifiedby RT-PCR.After it was cloned into T-vector and sequenced,pSG5 was used to transiently express peroxiredoxin I inIEC-6 by liposome-mediated transfection,and theexpression of peroxiredoxin I was evaluated by RT-PCRand Western blot.RESULTS:A DNA fragment about 750 bp was amplifiedfrom total RNAs of IEC-6 cells using specific primers ofperoxiredoxin I.The sequencing confirmed the codingregion was successfully cloned into T-vector,which wascompletely coinddent with the sequence in GeneBank.Afterthe EcoRI-BamHI fragment of T-vector containingperoxiredoxin I was inserted into pSG5,the recombinantplasmid was transferred to IEC-6 cells.RT-PCR assayshowed that a DNA fragment of 930 bp could be amplified,which indicated the transcription of pSG5-Prx.Western blotconfirmed the expression of peroxiredoxin I in IEC-6 cells.CONCLUSION:Mouse peroxiredoxin I can be successfullyexpressed in IEC-6 cells.
AIM: To clone and express mouse peroxiredoxin I in IEC-6 cells. METHODS: Total RNAs were isolated from cultured IEC-6 cells, and the coding region of peroxiredoxin I was amplified by RT-PCR. After it was cloned into T-vector and sequenced, pSG5 was used to transiently express peroxiredoxin I in IEC-6 by liposome-mediated transfection, and the expression of peroxiredoxin I was evaluated by RT-PCR and Western blot .RESULTS: A DNA fragment about 750 bp was amplified from total RNAs of IEC-6 cells using specific primers ofperoxiredoxin I. The confirmed confirmed that the coding region was successfully cloned into T-vector, which was completely linked to the sequence in GeneBank. AFterthe EcoRI-BamHI fragment of T-vector containing pesticide I was inserted into pSG5, the recombinant plasmid was transferred to IEC-6 cells.RT-PCR assays showed that a DNA fragment of 930 bp could be amplified, which shows the transcription of pSG5-Prx. Western blot confirmed the expression of peroxiredoxin I in IEC-6 cells. CONCLU SION: Mouse peroxiredoxin I can be successfullyexpressed in IEC-6 cells.