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公开(公告)号:KR1020010061173A
公开(公告)日:2001-07-07
申请号:KR1019990063661
申请日:1999-12-28
IPC: C12Q1/68
Abstract: PURPOSE: Provided is a vector containing PCR primers of tumor-suppressor gene and useful for the diagnosis of cancer, thereby the cancer can be easily diagnosed on a genetic level. CONSTITUTION: The vector TS-IS(KCTC 0708BP) contains primers of p14ARF, p16INK4A, p15INK4B, p21WAF1, p53 and pRb, and further contains HPRT primers, in which p14ARF, p16INK4A, p15INK4B, p21WAF1, p53 and pRb primers are produced by incubating liver cell lines of SK-Hep1, HepG2, HepG2.2.15, Hep3B, Chang liver, PLC/PRF5, SNU182, SNU354, SNU368, SNU387, SNU398, SNU423, SNU449 and SNU475 in an RPMI medium containing 10% of FBS; adding EDTA and trypsin into the fermented culture to remove the adhered cells; isolating total RNA using RNAzolTMB; and performing reverse transcription of the total RNA using a reverse transcriptase.
Abstract translation: 目的:提供含有肿瘤抑制基因的PCR引物的载体,可用于诊断癌症,从而可以在遗传水平上轻松诊断癌症。 构成:载体TS-IS(KCTC 0708BP)含有p14ARF,p16INK4A,p15INK4B,p21WAF1,p53和pRb的引物,并且还含有HPRT引物,其中通过孵育产生p14ARF,p16INK4A,p15INK4B,p21WAF1,p53和pRb引物 在含有10%FBS的RPMI培养基中的SK-Hep1,HepG2,HepG2.2.15,Hep3B,Chang肝,PLC / PRF5,SNU182,SNU354,SNU368,SNU387,SNU398,SNU423,SNU449和SNU475的肝细胞系; 向发酵培养物中加入EDTA和胰蛋白酶以除去粘附的细胞; 使用RNAzolTMB分离总RNA; 并使用逆转录酶进行总RNA的逆转录。
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公开(公告)号:KR100365482B1
公开(公告)日:2002-12-26
申请号:KR1019990063661
申请日:1999-12-28
IPC: C12Q1/68
Abstract: 본발명은종양억제유전자의발현을정량적으로분석함으로써암의진단에유용하게사용될수 있는진단용벡터에관한것으로, 간암을비롯한대부분의암에서자주불활성화를나타내는종양억제유전자인 p14, p16, p15, p21, p53 및 pRb의 PCR용각 프라이머쌍을포함하는본 발명의 TS-IS 벡터는, 경합 RT-PCR 방법을이용하여조직세포내 종양억제유전자들의발현유무와발현량을분석할수 있게해주기때문에, 암의발병여부를유전자수준에서진단할수 있을뿐 아니라, 어떤경로로암이발생했는지도예상할수 있어이후에진행될치료에많은정보를제공할수 있다.
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