Apoptosis induced by La( Sal)_2(Qu) in S. pombe with recombinant Fas gene
Li Xu
Abstract
Li Xu
Abstract
Objective To observe the apoptosis induced by La(C7H5O3) 2.(C9H6NO) in recombinant yeast.Meth-ods Total RNA of Jurkat cells was extracted by TRIZOL reagent.First-strand cDNA of Jurkat cells was synthesized by RT-PCR.The Fas genes were amplified by PCR and cloned into plasmid and formed a relevant shuttle carrier.The pREP3X-HA-Fas shuttle carriers were transformed into wild-type Schizosacharomyces pombe(S.pombe) by electroporation.The active recombination was identified by Western blotting and the apoptosis induced by La(C7H5O3) 2.(C9H6NO) was ob-served.Results The ratio of the bubble of cellular membrane,the fragmentation and caryolysis of cell nucleus induced by La(C7H5O3) 2.(C9H6NO) were 26.67%,22.86%,and 13.33%,respectively,detected with terminal deoxynucleotidyl transferase dUTP nick end labeling in treated group with significant differences compared with those of the control group (2.80%,3.74%,and 2.80%,respectively,P 0.01 for all).The apoptosis rate increased in treated group compared with that of the control group detected by flow cytometry(50.17% vs 1.24%,control group vs treated group,P 0.01).Conclusion La(C7H5O3) 2.(C9H6NO) could induce apoptosis by Fas in S.pombe.
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Objective To observe the apoptosis induced by La(C7H5O3) 2.(C9H6NO) in recombinant yeast.Meth-ods Total RNA of Jurkat cells was extracted by TRIZOL reagent.First-strand cDNA of Jurkat cells was synthesized by RT-PCR.The Fas genes were amplified by PCR and cloned into plasmid and formed a relevant shuttle carrier.The pREP3X-HA-Fas shuttle carriers were transformed into wild-type Schizosacharomyces pombe(S.pombe) by electroporation.The active recombination was identified by Western blotting and the apoptosis induced by La(C7H5O3) 2.(C9H6NO) was ob-served.Results The ratio of the bubble of cellular membrane,the fragmentation and caryolysis of cell nucleus induced by La(C7H5O3) 2.(C9H6NO) were 26.67%,22.86%,and 13.33%,respectively,detected with terminal deoxynucleotidyl transferase dUTP nick end labeling in treated group with significant differences compared with those of the control group (2.80%,3.74%,and 2.80%,respectively,P 0.01 for all).The apoptosis rate increased in treated group compared with that of the control group detected by flow cytometry(50.17% vs 1.24%,control group vs treated group,P 0.01).Conclusion La(C7H5O3) 2.(C9H6NO) could induce apoptosis by Fas in S.pombe.
Key concepts: Molecular biology, Apoptosis, DNA fragmentation, Terminal deoxynucleotidyl transferase, Jurkat cells, Recombinant DNA, Electroporation, Biology