中文摘要：

目的构建汉坦病毒S基因不同截短片段的重组表达质粒，并选择表达高活性的重组抗原应用于血清学诊断。方法从HTN型A537株基因组中扩增出S全长基因，构建TA-A537S克隆；以此为模板扩增出不同长度的截短的S片段，定向插入表达载体pET-30a．获得含不同片段的重组表达载体（pET-1-112．pET-1-119，pET-1-137，pET-1-200．pET-1-292，pET-1-316，pET-1-429，pET-6-119，pET-6-137，pET-6-200．pET-6-292．pET-6-405，pET-16-112，pET-16-137，pET-26-137），并在大肠杆菌BL21（DE3）中分别进行了表达；应用SDS-PAGE观察重组蛋白表达情况，应用Western blot分析重组抗原的活性，采用电洗脱、亲和层析等方法纯化重组蛋白．用间接ELISA法对重组蛋白的抗原性及抗原特异性进行检测。结果成功构建了含HVS全基因的TA-A537S克隆质粒；以截短的S基因片段构建的重组表达质粒中，既有表达又有活性的有pET-1-112，pET-1-119，pET-1-200，pET-1-292，pET-1-429，pET-6-119，pET-6-200，pET-6-292，pET-6-405；其中pET-6-119表达量最高且活性强；经电洗脱或Ni-NTA亲和层析可获得高纯度的重组蛋白e6-119；纯化的e6-119重组蛋白应用于ELISA检测疑似HFRS和不明原因的发热病人血清IgG，与IFAT比较，其敏感性和特异性分别为94．7％，95．6％。结论pET-6-119表达量高，易于纯化，并且具有良好的抗原性，是一种廉价、安全、可靠的诊断抗原。

英文摘要：

To construct the recombinant expression vector containing the different truncated genes encoding the nuclcocapsid protein of Hantavirus A537 in order to select the recombinant antigens with high expression capacity and higher antigenicity used for serological diagnosis, the full-length of S gene was amplified from Hantavirus strain A537 by PCR and inserted to expression vector pET-30a. From the recombinant plasmid TA-A537-S, several truncated genes from S gene of Hantavirus with different lengths were amplified and subcloned into the expression vector pET-30. The recombinant expression vectors con- structed were pET-1-112, pET-1-119, pET-1-137, pET-1-200, pET-1-292, pET-1-316, pET-1-429, pET-6-119, pET-6-137, pET-6-200, pET-6-292, pET-6-405, pET-16-112, pET-16-137, and pET-26-137. These gene fragments could be expressed respectively in E. coli BL21 （DE3） cells induced with IPTG. The expressed recombinant proteins were to be analyzed by SDS- PAGE and Western blotting and purified by electroelution or Ni-NTA affinity chromatographjy. The purified recombinant protein was used as antigen in an indirect ELISA to detect its antigenicity and specificity. It is demonstrated that TA-A537-S plasmid containing the full-length of S gene from Hantavirus was successfully constructed, in which several recombinant plasmids, i.e. pET-1-112, pET-1-119, pET-1-200, pET-1-293, pET-1-429, pET-6-119, pET-6-200, pET-6-292 and pET-6-405 were both expressed and active , among which pET-6-119 showed the highest amount of expression and excellent antigenicity. Satis- factory purity of the recombinant protein e6-119 could be achieved by electroeluton or Ni-NTA affinity chromatography. An indirect ELISA employing the purified e6-119 protein as coating antigen could detect the presence of specific IgG antibodies in sera of suspect patients with HFRS and fevers of unknown origin , with a 94.7% of sensitivity and a 95.60% of specificity, in comparison with those of IFAT.Therefore, the purified e6-119 protein may be a safe, cheap, sensitive a