| 1. | Someone tried this way but the yield is low . francis expressed truncated mutant pokeweed antiviral protein gene in pichia pastoris
目前已经有人在这方面做出了尝试，但蛋白表达产量还不高。 |
| 2. | Many studies revealed that pokeweed antiviral protein ( pap ) can control plant virus effectively
许多研究发现美洲商陆抗病毒蛋白( pokeweedantiviralprotein ， pap )对植物病毒具有良好的控制效果。 |
| 3. | To find out the difference between mut + and muts recombinants we compared their expression of pokeweed antiviral protein in the same conditions
在相同的培养条件下比较了mut ~ +和mut ~ s重组菌株表达pap的异同。 |
| 4. | Linearized the expression vector ppic9k - p including the truncated mutant pokeweed antiviral protein ( pap ) gene by restriction endonuclease sal i and transformed it electrically into pichia pastoris gs115 . mut + recombinants were selected by pcr and high yield mut + recombinant was picked out by double film immunoblotting
本研究将含有n末端信号肽和c末端毒性区缺失的pap基因的表达载体ppic9k - p用sali酶切线性化后，通过电击转化整合p . pastorisgs115菌株细胞中。 |
| 5. | The total rna was isolated from pokeweed ( phytolacca americana ) leaves using the method of guanidine isothiocyante and used as template to amplify the total length and deleted mutant pokeweed antiviral protein ( pap ) gene by rt - pcr and then the pap gene was cloned into pgem - t vector . the sequencing results showed that pap gene had 99 . 9 % identity comparing with the pap gene nucleotide sequence reported by lin et al ( 1991 ) . the iptg - inducible expression vector containing the pap gene was constructed and transferred into e . coli bl21 ( de3 ) - plyss
将缺失型pap基因克隆到植物表达载体pbi121中，通过液氮冷冻法将重组质粒转入农杆菌lba4404细胞中，然后采用叶盘法，在该农杆菌的介导下将pap基因导入普通烟草中，经过卡那霉素抗性筛选，最后获得了转pap基因的工程烟草植株，摩擦接种烟草花叶病毒( tmv ) ，与非转基因烟草相比，能够推迟症状表现达2月之久，说明pap基因能够在其它植物体内产生有活性的高抗病毒的蛋白质。 |
| 6. | Sds - page results showed that as to mut + recombinant highest yield was obtained after 4 days inducing and with the culture time prolonged it reduced . pokeweed antiviral protein gene expressed well when methanol concentration reached 10g / l . pokeweed antiviral protein obtained high yield in thin acidic culture medium ( ph6 . 0 - 6 . 4 ) and its quantity in total mass of secrete protein exceeded 30 %
Sds - page分析结果表明， mut ~ +组菌株在甲醇诱导第四天后pap在培养液中积累量达到最高水平，延长培养时间会导致产量下降；在10g / l的甲醇浓度诱导下， pap的表达量达到最高；培养基ph值在偏酸性条件下( 6 . 0 - 6 . 4 ) pap的表达量都维持在较高的水平。 |
