Title: Isolation of Genes Related with IL-6 Effect by a Rapid and Simple RT-PCR Method
Abstract: The significant results are gene expression when cytokine acts on receptors. To study genes related with IL 6 effect, here a simple and rapid PCR based protocol is utilized to detect and isolate cDNA fragments of the differential expressed genes between the IL 6 treated and untreated Sko007 cells. To generate cDNAs from most internal regions of mRNAs by the reverse transcription using three fully degenerated 6 mer oligonucleotide as primers. And then PCR amplification of the cDNAs with two or three longer primers that are arbitrary but defined sequences is carried on, The PCR amplification was repeated on the same cDNA templates (first step) with different sets of primers. PCR products were electrophoresed in the 2% agarose gel and then the recovered different fragments were used directly in cloning and sequencing. 3 different ESTs were obtained, one of which is a novel gene fragment, and its were proved to be IL 6 related genes by reverse Northern blot analysis. Further bioinformatics analysis indicated that new STRF17 fragments are expressed in many tissues and cells.
Publication Year: 2001
Publication Date: 2001-01-01
Language: en
Type: article
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Cited By Count: 3
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