A cDNA library was successfully constructed from Anip973, a human lung adenocarcinoma cell line with high metastatic potential. NIH3T3 cells were stably transfected using this cDNA library and screened for morphologic...A cDNA library was successfully constructed from Anip973, a human lung adenocarcinoma cell line with high metastatic potential. NIH3T3 cells were stably transfected using this cDNA library and screened for morphological changes in a soft agar assay. Genomic DNA was isolated from putative clones and the integrated sequence was retrieved by PCR and sequencing. Three known genes, ribosomal protein L23, hypothetical protein FLJ22104, and serine protease inhibitor, kazal type 6 and a number of 5＇-terminally truncated sequences were identified. Furthermore, cells transfected with ribosomal protein L23 was highly invasive compared with the empty vector as control （P 〈 0.02）. These results indicate that the expression cloning of cDNA libraries in NIH3T3 cells and subsequent screening for loss of contact inhibition in soft agar is a viable tool for identifying tumor-related genes and ribosomal protein L23 gene plays a role in cell movement and metastasis.展开更多
A full length cDNA named HongrESl was isolated and cloned by screening rat epididymis cDNA library using a mouse EST as a probe and 5'RACE followed. It contained 1590bp nucleotides and its predicted protein had 41...A full length cDNA named HongrESl was isolated and cloned by screening rat epididymis cDNA library using a mouse EST as a probe and 5'RACE followed. It contained 1590bp nucleotides and its predicted protein had 415 amino acid residues including a serpin (serine protease inhibitor) conserved domain. Tissue distribution pattern showed it was specifically expressed in adult rat epididymis; moreover, in situ hybridyza-tion indicated this gene was expressed in a limited region of the cauda epididymis near vas deference. Such kind of expression pattern sugested that HongrESl had potential function in male reproduction.展开更多
体内诱导基因是病原菌在宿主体内能够表达而体外培养时却不能表达的功能基因,其对病原体在宿主体内的生存和致病具有重要意义。体内诱导抗原技术(in vivo induced antigen technology,IVIAT)已广泛应用于筛选病原体体内诱导基因,相较于...体内诱导基因是病原菌在宿主体内能够表达而体外培养时却不能表达的功能基因,其对病原体在宿主体内的生存和致病具有重要意义。体内诱导抗原技术(in vivo induced antigen technology,IVIAT)已广泛应用于筛选病原体体内诱导基因,相较于其他用于筛选体内诱导基因的技术,IVIAT具有无需动物模型、检测病原菌在不同感染阶段产生的抗原等独特优势。IVIAT鉴定出的体内诱导抗原对病原体在宿主中的毒力、代谢及存活具有重要意义。现就IVIAT的原理、IVIAT筛选出的人类疾病相关病原菌的体内诱导抗原及其功能研究等作一概述。展开更多
基金This work was supported by Returning Scholars Fund of Heilongjiang Province (No. LC04C02) the Department of Education Overseas Researcher Fund of Heilongjiang Province (No. 1054HZ013).
文摘A cDNA library was successfully constructed from Anip973, a human lung adenocarcinoma cell line with high metastatic potential. NIH3T3 cells were stably transfected using this cDNA library and screened for morphological changes in a soft agar assay. Genomic DNA was isolated from putative clones and the integrated sequence was retrieved by PCR and sequencing. Three known genes, ribosomal protein L23, hypothetical protein FLJ22104, and serine protease inhibitor, kazal type 6 and a number of 5＇-terminally truncated sequences were identified. Furthermore, cells transfected with ribosomal protein L23 was highly invasive compared with the empty vector as control （P 〈 0.02）. These results indicate that the expression cloning of cDNA libraries in NIH3T3 cells and subsequent screening for loss of contact inhibition in soft agar is a viable tool for identifying tumor-related genes and ribosomal protein L23 gene plays a role in cell movement and metastasis.
基金supported by the Special Funds for Major State Basic Research Project(“973”No.G1999055901)State 863 High Technology R and D Project of China(No.2001AA221211)+3 种基金Shanghai Development Foundation of Science and Technology(No.01JC14046)Director foundation of Chinese Academy of Sciences,National Natural Science Foundation(No.30100064&39893320)Special funds of Biological Science and Technology from Chinese Academy of Science,NIH Forgarty International Center Grant 1-R03-TW01490-01A,CFDA No.93.934CAS knowledge creative program(KSCX2-SW-201).
文摘A full length cDNA named HongrESl was isolated and cloned by screening rat epididymis cDNA library using a mouse EST as a probe and 5'RACE followed. It contained 1590bp nucleotides and its predicted protein had 415 amino acid residues including a serpin (serine protease inhibitor) conserved domain. Tissue distribution pattern showed it was specifically expressed in adult rat epididymis; moreover, in situ hybridyza-tion indicated this gene was expressed in a limited region of the cauda epididymis near vas deference. Such kind of expression pattern sugested that HongrESl had potential function in male reproduction.
文摘体内诱导基因是病原菌在宿主体内能够表达而体外培养时却不能表达的功能基因,其对病原体在宿主体内的生存和致病具有重要意义。体内诱导抗原技术(in vivo induced antigen technology,IVIAT)已广泛应用于筛选病原体体内诱导基因,相较于其他用于筛选体内诱导基因的技术,IVIAT具有无需动物模型、检测病原菌在不同感染阶段产生的抗原等独特优势。IVIAT鉴定出的体内诱导抗原对病原体在宿主中的毒力、代谢及存活具有重要意义。现就IVIAT的原理、IVIAT筛选出的人类疾病相关病原菌的体内诱导抗原及其功能研究等作一概述。
