| 1. | Studies on virus resistance of transgenic tobacco with tmv replicase gene
转烟草花叶病毒复制酶基因烟草的病毒抗性研究
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| 2. | A novel rip named gynostemmin ( 27 kda ) was purified from the leaves and stems of gpentaphyllum
它具有很高的抗tmv活性。其分子量约为27kda 。
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| 3. | It showed excellent anti - tmv activity . and the n - terminal 19 amino acid residues are " dinfslagadgqtyn tfia " ( accession number p83206 in swiss - prot )
测得其n -端19个氨基酸序列： dinfslagadgqtyntfia ，与其它植物rip的同源性为10 73 ，与葫芦科rip的同源性为37 73 。
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| 4. | The resuh twed that mb designed in consisten the the complndny sequence of a sechon of the conservative regions of tmv - rna was suitable for the direet assay of tmv - rna . another contrl experiined had been pefformed to testify that our resuit was re1iable or not
Of 2inm之间有线性响应，检测下限达8pm ， t ： ；与共价交联的固定方法相比，此传感器具有较长的寿命和较好的稳定性。
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| 5. | The native expressed product from e . coli bl21 ( de3 ) strain , however , showed weak activity against tmv . gp609 and zwemu were inserted into pemu - mcs - n , an expression vector for monocotyledon . and rhxjb was inserted into pkylx71 : 35s2
将dna一8001连接到pet一sa表达载体上，在大肠杆菌blzi ( de3 )菌株中实现天然表达，表达产物对tmv的抑制效果很差，其原因可能是c一末端延伸序列的存在抑制了抗tmv活性的发挥。
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| 6. | Degenerate oligonucleotides to highly conserved regions of cucumis melo 1 - aminocyclopropane - 1 - carboxylic acid ( acc ) oxidase gene were used to prime the amplification of fragment of 128bp by ploymerase chain reaction ( pcr ) in samples of genomic dna from fruit of cucumis melo l . cv hetao flesh , which was cloned into plasmid vector pmd - 18 - t . the clon of antisense orientation were selected , and it was inserted downstream of camv35s promoter and enhancer " " of tmv into the plant expression vector pbinyxw , antisence expression vector pbinya was constructed . at the base that pollination and fertilization of cucumis melo l . cv hetao was studied , using pollen tube pathway transformate cucumis melo l . cv hetao , 76 fruit had been obtained , moreover , hardness and content of sugar were analysed
本实验以河套蜜瓜果肉基因组dna为模板，用甜瓜acc氧化酶基因特异寡核苷酸链为引物进行pcr扩增，得到128bp的扩增产物。将得到的扩增产物克隆到质粒载体pmd - 18 - t上，筛选反向克隆，然后将其反向构建到植物表达载体pbinyxw的camv35s启动子和tmv增强子“ ”的下游，构建成反义表达载体pbinya 。并在对河套蜜瓜授粉受精生物学研究的基础上，通过花粉管通道法转化河套蜜瓜，共获76颗瓜，并进行了硬度和含糖量的分析。
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| 7. | The deleted mutant pap gene was also cloned into yeast secreted expression ppic9k vector to form ppic9k ~ 3 , then the vector was transferred into pachia pastoris gs115 strain . the specific expression protein was secreted into the medium after inducing with methanol and the protein amount reached about 50 - 60 u g per millilitre measured by uv - absorbed methods in the supernatant of the medium via high density fermentation . sds - page results showed that there was one protein band in the gel which molecular weight was about 34ku
将缺失型pap基因克隆于酵母分泌型表达载体ppicgk构成重组载体，然后导入毕赤酵母（ p8chianastoris ）菌株gslls细胞中，在甲醇的诱导下，经过酵母高密度发酵进行pap的表达，经sds page分析，结果表明，在培养基上清液中含有一明显的特异性蛋臼条带，大小为34ku ，经western blotting分析，该蛋白与法国pap抗血清有特异性反应，体外活性检测表明该蛋白对tmv的侵染性具有高度的抑制性，说明该pap基因在毕赤酵母gs中也得到了正确表达。
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| 8. | The loop sequence of mb1 and mb2 were the anti sense and sense sequence ofing1 , respectively the sequence of mb3 was a piece of ssrna sequence in tobacco mosaic virus , which had no analogical to human gene . mbl was the most suitable probe because mbl had the highest fluorescence enhancemen after hybridizing wtth rna extrated froin normal cell
第三章，根据一种常见的病毒烟草花叶病毒( tmv )的核酸序列设计了分子信标荧光探针，由于tmv的遗传物质是rna ，分子信标又具有很高的特异性和灵敏度，因此感染了病毒粒子的植物叶片在经过简单处理后，可用分子信标检测叶片上。
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| 9. | Sds - page results showed that there was a clear target protein band in mut + recombinant supernatant after 48 hours of culturing , while a faint band only in muts recombinant after 72 hours . western - blotting result showed that there was no remarkable difference of yield between mut + and muts recombinants after 6 days induced . anti - virus activity tests revealed that culture supernatants of mut + and muts recombinants could inhibit tmv infection with high efficiency in the same concentration and there was no significant difference between them
结果表明，诱导培养48小时后， mut ~ +重组菌株表达产物在sds - page胶上显现出清晰的目的蛋白带，而mut ~ s重组菌株培养72小时才能显示微弱的目的带； western - blotting杂交信号强度表明，同样培养6天的mut ~ +和mut ~ s重组菌株表达产物在表达量上没有明显差别。
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| 10. | Both lines showed resistance to tomv - 0 , tomv - 2 , tomv - 1 , tmv - u , tmv - cg and susceptible to tomv - 2a . this result indicated that tm - 22 was functionally expression in receptor cell with distinct genetic background . virus specificity and gene express can function in distinct genus within a family
为了了解tm - 2 ~ 2基因的特性、功能及抗病机制，研究工作从三个方面进行，首先将tm - 2 ~ 2基因转化同科异属的烟草sr1 ，以确定tm - 2 ~ 2基因在模式植物的表达的可能性和功能的完整性。
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