非小细胞肺癌T7噬菌体展示文库的构建

来源 :中德临床肿瘤学杂志(英文版) | 被引量 : 0次 | 上传用户:tianwang782
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
Objective: Currently, only a limited numbers of tumor markers for non small lung cancer (NSCLC) diagnosis, new biomarker, such as serum autoantibodies may improve the early detection of lung cancer. Our objective is construction human lung squamous carcinoma and adenocercinoma T7 phage display cDNA library from the tissues of NSCLC patients. Methods: mRNA was isolated from a pool of total RNA extract from NSCLC tissues obtained from 5 adenocarcinomas and 5 squamous carcinomas, and then mRNA was reverse transcribed into double stranded cDNA. After digestion, the cDNA was inserted into T7Select 10-3 vector. The phage display cDNA library was constructed by package reaction in vitro and plate proliferation. Plaque assay and PCR were used to evaluate the library. Results: Two T7 phage display cDNA library were established. Plaque assay show the titer of lung squamas carcinoma library was 1.8×106 pfu, and the adenocarcinoma library was 5×106pfu. The phage titer of the amplified library were 3.2×1010 pfu/mL and 2.5 x 1010 pfu/mL. PCR amplifica-tion of random plaque show insert ratio were 100% (24/24) in adenocarcinoa library and 95.8% in human lung squamas carcinoma library (23/24). Insert range from 300 bp to 1 500 bp. Conclusion: Two phage display cDNA library from NSCLC were constructed.
其他文献
目的观察胰岛素样生长因子(IGF)2对Rh1肉瘤细胞生长活性和mTOR信号背景的影响。方法常规培养Rh1细胞,均用无血清培养液消除内源性因子影响后再用IGF-2(终浓度为10 ng/ml)刺激
NOB1基因是新发现的一个与细胞周期及转录调节有密切关系的基因。NOB1编码的蛋白质包括一个PIN(PilTamino terminus)结构域和一个ZNRD1(锌带)结构域,PIN结构域和转录相关,而Z
人工心脏瓣膜-患者不匹配是指植入瓣膜的有效开口面积相对于患者体型过小,术后残留明显的跨瓣压差进而影响患者预后.目前认为术后经胸超声测得的瓣膜有效开口面积指数是唯一
目的 分析宫腔镜对老年女性子宫内膜病变的诊断意义.方法 2005年1月至2008年12月就诊因怀疑子宫内膜病变行宫腔镜检查的老年女性患者130例,其中无症状组51例,有症状组79例.宫
目的观察间歇充气加压对腹腔镜胃癌手术患者下肢深静脉血流动力学的影响,并探讨其可能机制。方法将腹腔镜胃癌手术患者随机分成使用间歇充气加压组(实验组)和不使用间歇充气
1病例资料患者,男,26岁,因“无明显诱因咳嗽5个月”入院。胸部X线:双肺多发结节影,初步诊断为转移灶可能。胸部MRI:两肺多发结节(图1A),其余未见异常。体格检查及实验室检查
1临床资料患者,女性,37岁,因“腹痛、乏力、纳差4个月,加重伴腹胀3个月”于2009年4月16日收住入院。2008年12月中旬曾因月经不调自行服用土三七数日(具体用量不详)。2009年1
目的 :评价彩色超声显像技术在测量肠易激综合征 (IBS)患者内脏血液动力学方面的价值。方法 :18例肠易激综合征患者和 2 0名正常对照组于早晨空腹及餐后半小时分别测量肠系膜
为探究吕家坨井田地质构造格局,根据钻孔勘探资料,采用分形理论和趋势面分析方法,研究了井田7
Patients with stage Ⅲ N2 non-small cell lung cancer (NSCLC) are more than one third of all NSCLC patients.The 5-year survival rate of them is approximately 15%