Cloning, construction of prokaryotic expression vector and expression ofEscherichia coli cytosine deaminase gene

Cytosine deaminase gene ofEscherichia coli strain H-30 was cloned, and its initiation codon of ‘GTG’ was mutated to ‘ATG’ by PCR. Prokaryotic recombinant expression vector pBV220-CD was constructed. Clone with high enzyme activity were selected by detecting their specific activity of cytosine deaminase. 5-FC(5-FC, 5-fluorocytosine) could induce the lethal toxicity to cells containing active CD gene. DNA sequence analysis indicated that there were 16 altered bases and 5 of them resulted in the alteration of amino acids in predicted peptide by comparing DNA sequence of the clone H-30-CD-11 with high enzyme activity with CD gene reported in Gene Bank.     

Construction of High Level Expression Cell Lines of Erythropoietin *

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利用初等行变换求极大无关组的一些条件

本文讨论利用初等行变换求行向量组的极大线性无关组的方法,澄清一些线性代数教学用书中存在的一种模糊认识,并给出修正后的方法。     

广义块Pick型矩阵和带导数的Nevanlinna-Pick矩阵插值问题

建立了广义块Pick型矩阵和块Toeplitz矩阵之间的一种等价关系,并将其用于求解一类带导数的Nevanlinna-Pick矩阵插值问题     

腺病毒载体介导胸苷激酶基因/ACV对大鼠脑胶质瘤的离体和活体杀伤作用

建立缺陷型重组腺病毒载体介导的胸苷激酶基因治疗系统（Adtk／ACV）,进行离休和活体治疗大鼠C6脑胶质瘤研究．将质粒pAdtk与pJM17共转染腺病毒包装细胞──293细胞,同源重组后制备纯化的Adtk并以PCR证实;Adtk对离休C6细胞的杀伤作用随着Adtk滴度和ACV剂量增加而增强,并有旁观者效应,而未转染C6细胞和AdLacZ转染C6细胞均未被杀伤;扫描电镜下见Adtk／ACV处理的细胞呈明显病理改变;ACV可有效杀伤Adtk／C6细胞．在大鼠脑立体定向仪引导下于右额叶接种2×106个C6细胞,分别于第3,6,8和10天原位注射Adtk,同时腹腔注射ACV（100mg／（kg·d-1））,3d组、6d组、8d组大鼠成活期在90d以上,组织学检查未发现肿瘤细胞．10d组成活期（28．5±4．6）d,而C6未治疗组和AdLacZ／ACV治疗组成活期分别为（1．8±3．1）d和（14．0±2．2）d．本文建立的Adtk／ACV在离体和活体水平对大鼠C6脑胶质瘤有强烈的杀伤作用,效果确实,应用方便,ACV价格便宜,有潜在临床应用价值．     

外源基因（bar）在转基因燕麦（AvenasativaL.）后代中的遗传与表达

通过微弹轰击获得的含有ｂａｒ基因的转基因燕麦植株自交产生转基因后代。利用ＰＣＲ方法检测ｂａｒ基因在后代中的传递情况。     

高炉专家系统知识表达方法

分析了高炉炉况知识的特征。对各种知识表达方法进行了具体的分析并给出了实例,提出了采用“产生式规则＋框架＋语义网＋神经网络＋过程”的综合知识表达方法。     

ISOLATION,CHARACTERIZATION AND EXPRESSION OF THE COMPLEMENTARY DNA FOR HUMAN TUMOR NECROSIS FACTOR (TNF-α)

A cDNA for human TNF-α (615bp) was isolated by means of polymerase chain reaction (PCR) using first strand cDNA from PMA-induced HL-60 cells as template. The result from sequencing the 615 bp cDNA fragment indicated that it corresponded to the entire sequence of mature human TNF coding region. Direct expression of mature human TNF was achieved using a plasmid pHT-1 constructed by ligation of the cDNA and a synthetic DNA. The IPTG-induced bacterial product (hTNF) showed cytotoxicity to mouse L-929 cells. The TNF activity was further identified by neutralization of a specific monoclonal antibody against human TNF-α. Approximately 80,000 units of activity were detected per ml of culture at A600=2.     

跳帧转换编码中的运动矢量修正

在跳帧转换编码中,利用前向向量法进行运动矢量合成可以得到较好的图像信噪比。本文提出了用HAVS快速搜索算法对合成的运动矢量进行修正的方法,同时给出了一个最佳搜索窗口。实验结果表明,在±2像素内,用HAVS算法对合成的运动矢量修正,既能明显提高图像的信噪比,同时又能最大限度的降低计算量。     

血小板反应素-1活性片段的克隆与表达

选择两段血小板反应素（TSP-1）中抑制血管再生的活性片段，设计两对引物，使用RT-PCR的方法从人血细胞中进行克隆并对获得片段进行测序验证．克隆片段大小分别为723和522bp，命名为聪P-1-1和聪P-1-2．利用原核表达载体pET-29a获得大肠杆菌重组子，重组子经过IPTG诱导以包涵体的形式表达相应的多肽片段．再对包涵体进行体外溶解、纯化，得到了目的多肽．