K88菌毛基因的克隆及ST表位序列在菌毛基因上的融合

[目的]为了建立用于细胞表面展示的载体系统。[方法]利用PCR技术,以大肠杆菌质粒为模板扩增K88菌毛操纵子的结构基因faeC^faeH,并克隆到pBR322质粒载体中。合成一段替换序列,在菌毛抗原基因的超变区引入酶切位点ApaI和NcoI,合成耐热肠度素STII表位编码序列,并引入菌毛抗原基因的超变区。[结果]经PCR鉴定和限制性内切酶酶切证明,重组质粒pBR-fae插入片断大小为6.6kb,与预期相符。核苷酸序列分析证明,所得faeC^faeH序列正确。PCR筛选和测序验证证明,构建了K88菌毛-耐热肠度素STII的融合基因。[结论]试验成功获得了重组质粒pBR-fae-ST。

Characterization of ST13 Protein Expression in Human Colorectal Cancer Tissues

Objective: To characterize the expression of ST13 protein in human tissuesfor investigation of the function of colorectal cancer related gene ST13. Methods: ST13 ORF wascloned and over-expressed in E.coli. The recombinant ST13 protein was purified by affinitychromatography. ST13 monoclonal antibodies were generated and affinity purified with the recombinantprotein. Immunoblot and immunohistochemical staining were employed to analyze ST13 proteinexpression in human tissues. Results: The expression and purification of the recombinant ST13protein were confirmed by SDS-PAGE. The protein yield reached about 2.5 mg/L of induced bacterialculture with a purity of 91.3%. Three strains of hybridoma were obtained with antibody titers from10~4 to 10~5 in ascites fluids and with high specificity for ST13 protein. Immunoblot showed thatthe apparent Mr of ST13 protein in SW480 cells and human tissues estimated by SDS-PAGE mobility wasapproximately 50 000, which was about 10 000 larger than the 41 324 calculated, but theglycosylation of the protein was excluded. Computer modeling revealed the protein to be ahydrophilic molecule. Immunohistochemical staining showed that ST13 protein was evenly distributedin cytoplasm and expressed in colon, stomach, liver, and other epithelial cells. Differences in thestaining intensity of the protein were observed between normal and cancer tissues as well as amongdifferent normal or carcinoma tissues. Conclusion: ST13 protein is a cytoplasmic molecule with anapparent Mr of 50 000. The protein is expressed in colorectal and other epithelial tissues. Theexpression level of the protein is down-regulated in colorectal cancer and varies among differentnormal and/or carcinoma tissues. Comparison of cDNA sequences and protein characteristics indicatesthat ST13 protein and hsp70-interacting protein (Hip) are same proteins, raising the possibilitythat ST13 protein is involved in the development of colorectal cancer through Hsp70 molecularchaperone machinery.

Evolution of Surface Cold Patches in the North Yellow Sea Based on Satellite SST Data

Ten years （from 2005 to 2014） of satellite sea surface temperature （SST） data from the Advanced Very High Resolution Radiometer （AVHRR） are analyzed to reveal the monthly changes in surface cold patches （SCPs） in the main areas of the Northern Yellow Sea （NYS）. The Canny edge detection algorithm is used to identify the edges of the patches. The monthly changes are de- scribed in terms of location, temperature and area. The inter-annual variations, including changes in the location and area of the SCPs from 2010 to 2014, are briefly discussed. The formation mechanisms of the SCPs in different periods are systematically analyzed using both in situ data and numerical simulation. The results show that from May to October, the location and area of the SCPs re- main stable, with a north-south orientation. The SCPs altogether cover about I° of longitude （124°E-125°E） in width and 2° of lati- tude （37.5°N-39.5°N） in length. In November, the SCP separates from the Jangsan Cape and forms a closed, isolated, and approxi- mately circular cold patch in the central NYS. From May to October, the upweUing that leads to the formation of the SCP is mainly triggered by the headland residual current, wind field, climbing movement of the current and secondary circulation at the tide front. In November, cyclonic circulation in the NYS is primarily responsible for generating the upwelling that leads to the formation of the closed and isolated SCE

最近公共祖先算法在管道运输的应用

介绍最近公共祖先算法的原理和应用,并总结四种最近公共祖先算法:欧拉序结合ST表法、倍增法、并查集结合Tar⁃jan算法和树链剖分法,重点剖析欧拉序结合ST表法具体的算法过程。提出数列区间操作问题的解决方案,并将此推广在树中,讨论两种树链操作方式:树上差分和相对于根结点的树上标记法。将最近公共祖先算法与树上标记结合,并运用在管道运输问题上,能在O(n log2n)的算法复杂度下,监控管道运输网络的最大压力值。