Gene Expression and Activities Analysis of a New Fusion Protein (RGD)3/tTF

为了发展一种新型的融合蛋白(RGD)3/tTF用于肿瘤血管的选择性栓塞治疗,利用PCR技术重组(RGD)3/tTF融合基因,克隆于pET22b(+)载体,表达于E.coliBL21(DE3)。用镍柱纯化融合蛋白。凝血实验与FⅩ活化实验检测融合蛋白tTF组分的活性。间接ELISA分析(RGD)3/tTF与αvβ3的特异结合能力。pET22b(+)/(RGD)3/tTF重组质粒成功获得并表达于E.coliBL21(DE3)。纯化蛋白(RGD)3/tTF能有效诱发血液凝固,活化FⅩ。(RGD)3/tTF与αvβ3的特异结合能力比RGD/tTF提高了32%。新型融合蛋白(RGD)3/tTF已在E.coli系统成功表达,表达蛋白保持tTF的活性并显示比RGD/tTF更高的与αvβ3的结合能力。 【英文摘要】 To develop a new fusion protein (RGD)3/tTF for the therapy of the selective thrombosis of tumor blood vessels. The fused gene (RGD)3/tTF was reconstructed by PCR, was cloned into vector pET22 b(+),and expressed in E.coli BL21(DE_3). The fusion protein was purified through Nickel-affinity chromatography column. The tTF activity of the fusion protein was detected by clotting assay and FⅩ activation assay. The specific binding of (RGD)3/tTF to α_vβ_3 was analyzed by indirect ELISA. The recombinant plasmid pET2...福建省自然科学基金(No.C0410004);; 厦门大学科技创新基金资助(No.XDKJCX20053026)。~

Influence of p-GaN Annealing on Optical Properties of InGaN MQWs

近年来,n2退火和O2退火均被用于激活P-gAn中的Mg受主以提高P-gAn中的空穴浓度。基于两种退火技术,系统地研究了n2退火和O2退火对lEd样品电学性能及光学性能的影响。电流电压特性的测试结果显示,在较低温度(500℃)下O2退火就可以达到与n2高温退火(800℃)相似的电学特性。变温光致发光测试表明,n2高温退火会在IngAn量子阱中形成In团簇,In团簇作为深的势阱增加了对载流子的束缚,能够将载流子更好地局限在势阱中。然而In团簇形成的同时也伴随着大量位错的产生,使其IngAn量子阱中的位错密度大幅度提高,因此室温下n2退火样品的辐射复合效率低于O2退火样品的辐射复合效率。In recent years,thermal annealing in either N2 ambient or O2 ambient was used to activate the Mg-doped GaN epilayer and thus improve the density of holes in p-GaN.The electrical and optical properties of LED samples annealed in different ambient were systematically investigated.The test results of I-V characteristics show that samples annealed at low temperature(500 ℃) in O2 ambient and high temperature(800 ℃)in N2 ambient show similar current-voltage characteristics.The temperature-dependent photoluminescence(PL)measurement shows that high-temperature thermal annealing in N2 ambient can induce In clusters in InGaN multiple quantum well(MQWs).The deep traps induced by In clusters can work as localized centers which can enhance the confinement of carriers,the cavriers can be better boanded in well.However,there are much more dislocations out of the trap centers caused by high-temperature annealing,the dislocation density of InGaN MQWs increased significantly.Therefore,at room temperature,the radiative efficiency of the sample annealed in N2 ambient was lower than that annealed in O2 ambient.国家自然科学基金(10974165;91023048;61106044); 高等学校博士学科点专项科研基金(20110121110029

用于肿瘤血管靶向治疗的RGD/tTF融合蛋白的表达及活性鉴定

目的:制备用于肿瘤血管靶向治疗的重组RGD/tTF融合蛋白,并鉴定其生物学活性。方法:利用PCR技术构建RGD与tTF的融合基因,克隆至表达载体pET22b(+),在E.coliBL21(DE3)中表达,镍亲和层析柱纯化。凝血实验和FⅩ活化实验鉴定融合蛋白中tTF的活性,间接ELISA分析RGD活性。结果:获得序列正确的RGD/tTF/pET22b(+)重组子,融合蛋白在E.coliBL21(DE3)中高效表达。纯化后的融合蛋白具有活化FⅩ、引起血液凝固的功能,且能与αvβ3特异性结合。结论:成功构建RGD/tTF/pET22b(+)重组子,RGD/tTF融合蛋白具有TF活性且与αvβ3特异性结合,为进一步研究其体内特异性诱发肿瘤组织血管栓塞功能创造了条件

ヘイアンジョガクイン ニ オケル カンコウ ホスピタリティ キョウイク ノ セイリツシ オ メグル ケンキュウ

大学における観光系学部学科開設に伴い、観光ホスピタリティ教育という分野の研究が興隆した。しかし、同分野の研究は概念の定義や手法、女子教育との連関など諸課題を抱えている。本稿では、現代的女子教育の可能性の提示と、多様化に合致した観光ホスピタリティ教育の構築のために、女子教育を振り返り、観光ホスピタリティ教育との接点を検討する。構成ではまず観光学部設置の経緯を追い、平安女学院大学を事例に観光ホスピタリティ教育の現状を述べる。次に日本の女子教育に影響を与えた女子ミッションスクールの理念と、良妻賢母教育の関係に言及する。最後に、平安女学院の変遷から、女子教育が良妻賢母、英語、秘書教育から観光ホスピタリティ教育に移行した様とその連関を明らかにする。以上により、「良妻賢母教育」が現在の国内外観光客への「おもてなし」の原点として、「観光ホスピタリティ教育」に導入されたことへの裏付けが可能になった。The department of tourism and hospitality management has been founded in many colleges.Especially most of women\u27s colleges have established these courses so as to make students get employment in tourism industries. However, these researches and educational methods related to women\u27s education are still in progress. This paper insists on new possibilities for the development of tourism and hospitality courses in contemporary women\u27s education on undergraduate level. These educational systems could be expanded to respond to diverse needs from tourism industries. Thepaper also focuses on the historical background of women\u27s education related to tourism and hospitality. The paper discusses three main issues. First, it demonstrates the process of the establishment of atourism department at Heian Jyogakuin University and explains its present status. Second, it refers to the philosophy of girls\u27 Christian universities in Japan and its encouragement to make girls goodwives and wise mothers. Third, it demonstrates how Heian Jyogakuin University shifted its gears from making a good wife and wise mother into English and secretarial education and tourism and hospitality education. Finally the paper concludes that the education to make a good wife and wisemother was the starting point of tourism and hospitality education

人源化抗人肺癌单域抗体基因的构建、表达及活性分析

目的:构建人源化的抗人肺癌单域抗体hu3D3VH基因,在大肠杆菌中表达,对其蛋白活性进行分析。方法:采用CDR移植技术对mAb3D3的重链可变区进行人源化,通过重叠PCR获得hu3D3VH的基因。构建pET22(b+)/hu3D3VH表达载体,并转化大肠杆菌BL21(DE3),在IPTG诱导下表达。表达产物通过Ni亲和层析柱纯化。采用间接ELISA和竞争抑制ELISA法进行活性分析。结果:通过重叠PCR获得序列正确的目的基因。目的蛋白以包涵体的形式表达,表达量占菌体总蛋白的30%以上。纯化后,目的蛋白的纯度达95%以上。hu3D3VH具有与亲本抗体相同的抗原反应性,并能抑制mAb3D3与L342细胞的结合。结论:获得的人源化单域抗体hu3D3VH,保留了与mAb3D3相同的反应性和特异性,为进一步临床应用奠定了基础

诱骗受体DcR3的基因构建、表达及特异性鉴定

目的:构建适于原核表达的人诱骗受体DcR3基因,并进行重组蛋白的表达纯化及特异性鉴定。方法:查得人DcR3cDNA全序列,将其分段设计引物,通过重叠PCR获得DcR3基因。构建pET-22b(+)/DcR3表达载体,转化大肠杆菌Rosseta-gami,IPTG诱导表达,Ni柱纯化。采用ELISA进行特异性鉴定。结果:通过重叠PCR获得了编码正确氨基酸序列的目的基因。目的蛋白以包涵体的形式表达,表达量占菌体总蛋白的30%以上。纯化后,蛋白纯度达95%以上。ELISA结果表明所纯化的蛋白可与抗DcR3抗体发生特异结合。结论:诱骗受体DcR3基因的成功构建、表达及纯化,为进一步的功能研究奠定了基础