AIM: To study the effect of short hairpin RNAs (shRNAs)expressed from DNA vector on hTERT expression.METHODS: Oligonucleotides coding for four shRNAs against hTERT were cloned into a mammalian shRNA expressionvector p...AIM: To study the effect of short hairpin RNAs (shRNAs)expressed from DNA vector on hTERT expression.METHODS: Oligonucleotides coding for four shRNAs against hTERT were cloned into a mammalian shRNA expressionvector pUC18U6 to form pUC18U6ht1-4, which were thenintroduced into HepG2 cells by using liposome-mediated transfection. HepG2 cells transfected by pUC18U6 and pUC18U6GFPsir, which expressed shRNA against green fluorescent protein (GFP), were used as controls. hTERT mRNA in the transfected cells were quantified by using real-time fluorescent RT-PCR.RESULTS: Among the four shRNAs against hTERT, two decreased the hTERT mRNA level. Compared with the controls, pUC18U6ht which expressed the two shRNAs reduced hTERT mRNA by 39% and 49% (P＜0.05).CONCLUSION: hTERT expression is inhibited by the shRNAs expressed from the DNA vector.展开更多
To clone HPV16 E2 gene from a biopsied cervical cancer sample Materials & Methods HPV16 E2 gene was amplified from specimen derived from a HPV 16 positive patient, then cloned and sequenced. Results The full ...To clone HPV16 E2 gene from a biopsied cervical cancer sample Materials & Methods HPV16 E2 gene was amplified from specimen derived from a HPV 16 positive patient, then cloned and sequenced. Results The full length of HPV 16 E2 gene was successfully cloned. In comparison with the prototype accepted by GenBank, six point mutations in HPV 16 E2 nucleotide acid sequence were identified. Of them, three were missense, and one was in the overlapping E4 gene and was synonymous to E4. Conclusion HPV16 E2 gene was successfully cloned, and some nucleotide acids in its sequence were different from the prototype.展开更多
基金Supported by the Shaanxi Province Natural Science Foundation,No. 2003C217
文摘AIM: To study the effect of short hairpin RNAs (shRNAs)expressed from DNA vector on hTERT expression.METHODS: Oligonucleotides coding for four shRNAs against hTERT were cloned into a mammalian shRNA expressionvector pUC18U6 to form pUC18U6ht1-4, which were thenintroduced into HepG2 cells by using liposome-mediated transfection. HepG2 cells transfected by pUC18U6 and pUC18U6GFPsir, which expressed shRNA against green fluorescent protein (GFP), were used as controls. hTERT mRNA in the transfected cells were quantified by using real-time fluorescent RT-PCR.RESULTS: Among the four shRNAs against hTERT, two decreased the hTERT mRNA level. Compared with the controls, pUC18U6ht which expressed the two shRNAs reduced hTERT mRNA by 39% and 49% (P＜0.05).CONCLUSION: hTERT expression is inhibited by the shRNAs expressed from the DNA vector.
文摘To clone HPV16 E2 gene from a biopsied cervical cancer sample Materials & Methods HPV16 E2 gene was amplified from specimen derived from a HPV 16 positive patient, then cloned and sequenced. Results The full length of HPV 16 E2 gene was successfully cloned. In comparison with the prototype accepted by GenBank, six point mutations in HPV 16 E2 nucleotide acid sequence were identified. Of them, three were missense, and one was in the overlapping E4 gene and was synonymous to E4. Conclusion HPV16 E2 gene was successfully cloned, and some nucleotide acids in its sequence were different from the prototype.
