多重荧光PCR同时检测转基因成分35S和Nos方法的建立

根据商品化转基因作物中常用的花椰菜花叶病毒启动子 (CaMV 35S)和根癌农杆菌终止子 (Nos)的序列特点 ,设计并合成了两对引物和相对应的荧光双链探针 ,建立一种应用荧光双链探针的多重荧光PCR同时检测转基因成分 35S启动子和Nos终止子的方法 .并利用该方法对马铃薯、大豆、玉米、甜椒、番茄等 11份实物样品进行了检测 ,其中有 5份样品结果阳性 .结果表明所建立的多重荧光PCR方法能同时检测出 35S和Nos双组分 ,较常规PCR技术更为简便、快速、准确 ,有很好的应用前景

Expression of Fusion Protein of Parathyroid Hormone and Transferrin N-terminal Half-molecule in Pichia pastoris

利用重叠PCR技术将PTH(parathyroidhormone,甲状旁腺激素)基因与TFN(transferrinN_terminalhalf_molecule,转铁蛋白N端半分子)基因在体外融合,融合基因克隆至真核表达载体pPIC9中,转化毕赤酵母GS115。转化子经甲醇诱导后,融合蛋白得到了表达并分泌到发酵上清液中。经SPSepharoseFF阳离子交换层析、PhenylSepharoseFastFlow疏水层析纯化获得了纯度大于95%的PTH_TFN样品。Westernblot分析及腺苷酸环化酶实验证明融合蛋白中的PTH具有与抗PTH抗体结合能力及刺激腺苷酸环化酶的活性,铁饱和实验证明融合蛋白中的TFN和单独的TFN具有相同铁结合能力。因而TFN可望作为PTH的天然运输载体。The fused gene (PTH_TFN) of parathyroid hormone (PTH) gene and transferring N_terminal half_molecule (TFN) gene was amplified by multiple PCR and inserted into pPIC9 vector. The recombinant plasmid pPIC9_PTH_TFN was transformed into Pichia pastoris GS115 by PEG. After methanol induction, the target protein was expressed in fermentation supernatant at high level.The fused protein PTH_TFN with purity being higher than 95% was finally obtained after purification through two_step chromatography : SP Sepharose Fast Flow and Phenyl Sepharose Fast Flow.Western blot analysis and adenylate cyclase assay proved that the fused protein exhibited the bioactivity to stimulate cAMP synthesis and the ability to bind Fe ~3+ in the Fe ~3+ saturation study as the recombinant TFN did indicating that TFN could be used as the transcellar carrier of PTH.国家高技术研究与发展项目基金资助(No.2004AA215172)。~

中国海及邻近区域碳库与通量综合分析

中国海总面积约470万平方公里,纵跨热带、亚热带、温带、北温带等多个气候带.其中,南海北依\"世界第三极\"青藏高原、南邻\"全球气候引擎\"西太平洋暖池,东海拥有全球最宽的陆架之一,跨陆架物质运输显著,黄海是冷暖流交汇区域,渤海则是受人类活动高度影响的内湾浅海.中国海内有长江、黄河、珠江等大河输入,外邻全球两大西边界流之一的黑潮.这些鲜明的特色赋予了中国海碳储库和通量研究的典型代表意义.文章从不同海区（渤海、黄海、东海、南海）、不同界面（陆-海、海-气、水柱-沉积物、边缘海-大洋等）,以及不同生态系统（红树林、盐沼湿地、海草床、海藻养殖、珊瑚礁、水柱生态系统等）多层面对海洋碳库与通量进行了较系统地综合分析,初步估算了各个碳库的储量与不同碳库间的通量.就海气通量而言,渤海向大气中释放CO2约0.22Tg Ca-1,黄海吸收CO2约1.15Tg Ca-1,东海吸收CO2约6.92～23.30Tg Ca-1,南海释放CO2约13.86～33.60Tg Ca-1.如果仅考虑海-气界面的CO2交换,中国海总体上是大气CO2的\"源\",净释放量约6.01～9.33Tg Ca-1.这主要是由于河流输入以及邻近大洋输入所致.河流输入渤黄海、东海、南海的溶解无机碳（DIC）分别为5.04、14.60和40.14Tg Ca-1,而邻近大洋输入DIC更是高达144.81Tg Ca-1,远超中国海向大气释放的碳量.渤海、黄海、东海、南海的沉积有机碳通量分别为2.00、3.60、7.40、7.49Tg Ca-1.东海和南海向邻近大洋输送有机碳通量分别为15.25～36.70和43.39Tg Ca-1.就生态系统而言,中国沿海红树林、盐沼湿地、海草床有机碳埋藏通量为0.36Tg Ca-1,海草床溶解有机碳（DOC）输出通量为0.59Tg Ca-1;中国近海海藻养殖移出碳通量0.68Tg Ca-1,沉积和DOC释放通量分别为0.14和0.82Tg Ca-1.总计,中国海有机碳年输出通量为81.72～103.17Tg Ca-1.中国海的有机碳输出以DOC形式为主,东海向邻近大洋输出的DOC通量约15.00～35.00Tg Ca-1,南海输出约31.39Tg Ca-1.综上,尽管从海-气通量看中国海是大气CO2的\"源\",但考虑了河流、大洋输入、沉积输出以及微型生物碳泵（DOC转化输出）作用后,中国海是重要的储碳区.需要指出的是,文章数据是基于中国海各海区碳循环研究报道,鉴于不同研究方法上的差异,所得数据难免有一定的误差范围,亟待将来统一方法标准下的更多深入研究和分析.国家重点研发计划项目(编号:2016YFA0601400);;国家自然科学基金项目(批准号:91751207、91428308、41722603、41606153、41422603);;中央高校基础研究项目(编号:20720170107);;中海油项目(编号:CNOOC-KJ125FZDXM00TJ001-2014、CNOOCKJ125FZDXM00ZJ001-2014)资

A CASE OF ILEUS BY THE INFECTION OF TYPE X LARVAE OF THE SUBORDER SPIRURINA AFTER EATING RAW FIREFLY SQUIDS

A 53-year-old male was hospitalized on April 12th, 2002 because of an upper abdominal pain that started in the morning of April 11th and gradually worsened. Ileus was suspected based on an X-ray of the abdomen. A medical interview revealed that the patient had eaten raw firefly squids, 2 days before the hospitalization. The abdominal symptoms disappeared in a few days during which the patient was treated conservatively. A specimen from the patient's sera reacted with the esophageal glands in frozen sections of a type X larva of the suborder Spirurina. The infection of the type X larvae of unidentified splruroid nematodes from squids was considered to be the cause, because those larvae have recently been to reported as the cause of visceral and cutaneous. Although the infection rate in W. scintillans is relatively low, a risk of human infection with type X larvae should be taken into a consideration in the consumption of raw squids