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1.
小鼠B7—1cDNA克隆,测序,表达及其抗瘤效应的初步探讨   总被引:4,自引:0,他引:4  
用反转录PCR的方法,从BALB/c小鼠脾细胞中扩增出B7-1cDNA后,插入pcDNA3质粒中构建成小鼠B7-1cDNA的真核表达载体pCD-mB7.1,经酶切鉴定和序列分析证实此表达载体中插入的B7-1cDNA的序列与献报道一致。  相似文献   

2.
OA能诱发人神经母细胞瘤SK细胞进行编程死亡.死亡细胞缩小变圆,细胞质凝聚,DNA有控降解成约200bp左右的片段,且这一过程可由蛋白质合成抑制剂亚胺环己酮(CHX)抑制.将编码Bc1-2全长蛋白质的cDNA植入pXJ41neo载体中,使其表达由HCMV病毒启动子控制.形成的顺义(pBcl-2-S)及反义(pBcl-2-AS)表达质粒经转染导入SK细胞中获得稳定转染子,West-ern印迹表明顺义转染子表达较大量的26kdBc1-2蛋白,而反义转染子则不表达.增强表达的Bc1-2基因产物对OA引SK细胞编程死亡无抑制效应.  相似文献   

3.
小白菜苏云金牙孢杆菌CryIB基因的遗传转化   总被引:3,自引:0,他引:3  
用限制SphI和EcoRI消化质粒pBYTESp101-1和pBQX,将pBQX上的经发行过的苏云金芽直菌杀虫晶体蛋白基因CryIB取代pBYTESp101-1上的GUS基因,得到一个中间克隆pBIWIQ-1。用限制酶HindⅢ消化质粒pBIWIQ-1,回收含CryIB基因的5.1kbDNA乍段,插入质粒pBIN19的Hind0283位点,构建成具卡那霉素抗性的植物表达载体pBIWIQ-5.ET  相似文献   

4.
将克隆入pGEM7zf(+)的甜菜坏死黄脉病毒(BNYVV)新疆分离物外壳蛋白(CP)基因的cDNA的pGEB3,用XbaI切下,Klenow补平,再用BamHI切下cDNA片段。用NdeI切开原核表达载体pJW2,用Klenow补平,再用BamHI切去小片段。将该cDNA与pJW2大片段用T4连接酶连接,构建了BNYVVCP基因的表达载体pJWB4,转化到大肠杆菌DH5α。经培养和高温诱导,pJWB4成功地表达出了BNYVV的外壳蛋白。将pGEB3和pBI121用Xbal和BamHI酶切T4连接酶连接,构建了BYVVCP基因的植物表达载体,转化到DH5α,筛选出正向连结的阳性克隆pBIB3,转化入农杆菌LBA4404(pAL4404),经用PCR扩增和γ32P标记的探针杂交约证实为阳性克隆。往甜菜植株中转化工作正在进行中。  相似文献   

5.
RATS1基因对人食管癌细胞c—myc基因表达的降调…   总被引:1,自引:0,他引:1  
将含有RATS1cDNA的真核表达载体pCDV1与含有neo抗性基因的表达载体pDOR-neo经Lipofectin法共转染人食管癌细胞系EC109细胞。结果发现:转染RATS1基因后的抗性克隆细胞生长受到抑制,形态发生明显改变,并且逐渐脱落死亡。经RT-PCR分析发现,转染后有RATS1基因表达的细胞克隆,c-myc基因表达降低,两者呈反向相关,即RATS1基因表达愈高,c-myc基因表达降低愈  相似文献   

6.
丙型肝炎病毒核酸疫苗的研究   总被引:1,自引:0,他引:1  
从丙型肝炎病人的阳性血清中提取出丙型肝炎病毒RNA,通过RT-PCR的方法获得丙型肝炎病毒C区基因片段。并将此片段克隆到真核细胞表达载体pcDNA3.1(+),得到重组质粒pcDNA-HCV/C,再将其通过肌肉注射免疫BALB/小鼠后,小鼠产生了抗HCV/C区抗体。  相似文献   

7.
HSV1-tk基因的部分DNA序列分析   总被引:3,自引:0,他引:3  
采用PCR技术从商品质粒pHSV106扩增出HSV1-tk基因(1128bp),PCR产物用BamHI和EcoRI双酶切后定向克隆至真核表达载体pcDNA3,然后以重组质粒pcTK为模板tk基因的5端引物为DNA测序引物进行部分DNA序列分析,部分DNA序列分析证明PCR产物为HSV1-tk基因正确无误,成功筛选到HSV1-tk基因的真核表达载体pcTK,并为利用tk基因在实验动物体内进行自杀性基  相似文献   

8.
牛酪蛋白cDNA基因克隆的改建   总被引:1,自引:1,他引:0  
牛酪蛋白全长cDNA克隆pBaS1c184X GLⅢ和COrI双酶切,回收基中的酪蛋白基因编码片段,与经BamHI,SmaI双酶切的植物表达载体pBI12.12分两步连接;第一步载体的BamHI末端与酪蛋白基因的BglⅡ粘性末端连接;第二步用T4DNA多聚酶补平酪蛋白基因的EcoTRⅠ末端,再与载体SmaⅠ进行平末端连接而环化。  相似文献   

9.
将插入pBluescribe的人红细胞生成素受体cDNA,用EcoRI和BamHI酶解,分离得到的基因片段,插入到经EcoRI和RamHI消解的表达载体pGEX-3X中,得到了重组表达质粒pGEX-3X/HEPOR。生组质粒pGEX-3X/hEPOR含有tqc启动子,EPOR膜外结构域基因5’端与谷胱甘肽转移酶编码基因融合,阳性重组子在大肠村菌中经IPTG诱导表达GST-hEPOR,重组表达菌裂解  相似文献   

10.
从蓝藻Calothrixsp.PCC7601染色体DNA中分离出含藻蓝蛋白基因cpcB2A2的略3.8KbDNA片段,与质粒pUC18重组,构建成一种重组质粒,pPC218-PCZ,转化E.codiJM109,获得了一系列克隆子,经斑点分子杂交和限制性内切核酸酶分析,筛选出了初步认为含有cpcB2A2基因的克隆子。  相似文献   

11.
Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.  相似文献   

12.
The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.  相似文献   

13.
王慧 《科技信息》2008,(10):240-240
Wuthering Heights, Emily Bronte's only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.  相似文献   

14.
The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3~4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰~ -7.8‰, PDB) and matrix dolomites (-6.6‰~ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084~0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078~0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.  相似文献   

15.
何延凌 《科技信息》2008,(4):258-258
Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women's language from these perspectives: pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women's language, more attention should be paid to language use in social context. What's more, the linguistic phenomena in a speaking community can be understood more thoroughly.  相似文献   

16.
Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.  相似文献   

17.
以AC-13级配为基础,将橡胶颗粒代替部分集料掺入混合料中,以低温弯曲试验为评价方法对不同橡胶颗粒掺量下沥青混合料的低温抗裂性进行研究,并引入应变能密度值对混合料的低温抗裂性进行综合评价.试验结果表明:橡胶颗粒沥青混合料试件的破坏微应变均超过2 300,满足冬寒区的技术指标;无论是否掺加橡胶颗粒,随着温度的下降,沥青混合料破坏时的最大弯拉强度增大,弯拉应变降低,劲度模量增大;弯曲应变能密度在胶粒掺量为1%左右时具有较大的弯曲应变能密度值,此时橡胶颗粒沥青混合料具有较好的低温抗裂性.  相似文献   

18.
AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…  相似文献   

19.
理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。  相似文献   

20.
As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.  相似文献   

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