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Article in Chinese | WPRIM (Western Pacific) | ID: wpr-248534

ABSTRACT

<p><b>OBJECTIVE</b>To establish a real-time quantitative RT-PCR method for the detection of expression of bcl-2 mRNA.</p><p><b>METHODS</b>The vector containing bcl-2 gene as standard template was constructed with T-A cloning technique. The fluorogenic probe (i.e.,Taq man probe) was used to establish a real-time RT-PCR. bcl-2 mRNA expression level in Burkitt's lymphoma pre-and post-treated with bcl-2 antisense phosphothioate oligodeoxynucleotides (AS-PS-ODN) was determined with real-time quantitative RT-PCR, also with semi-quantitative RT-PCR.</p><p><b>RESULTS</b>The expression of bcl-2 mRNA in Burkitt's lymphoma treated with bcl-2 AS- PS-ODN decreased significantly and no changes of bcl-2-mRNA expression in group treated with nonsense ODN were noticed. The semi-quantitative RT-PCR results also demonstrated that bcl-2 level varied as detected with real-time fluorogenic quantitative RT-PCR, but less sensitive and accurate.</p><p><b>CONCLUSION</b>Detection of bcl-2 mRNA expression with the fluorogenic real-time quantitative RT-PCR is more accurate and sensitive than semi-quantitive RT-PCR.</p>


Subject(s)
Humans , Electrophoresis, Agar Gel , Methods , Gene Expression , Proto-Oncogene Proteins c-bcl-2 , Genetics , RNA, Messenger , Reverse Transcriptase Polymerase Chain Reaction
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