美洲商陆

The pokeberry belongs to perennial herb and it is one of manganese in hyperaccumulation plants.

美洲商陆属于商陆科多年生草本植物，是一种适应性强的锰超积累植物。

Type genus of Phytolaccaceae: pokeweed.

商陆科的模式属；美洲商陆。

Pokeweed antiviral protein belongs to Ribosome-inactive Proteins . PAP has potent antiviral activity against many plant virus. pTI was used as the template to amplify the deleted mutant PAP gene (N-deleted and N-, C-deleted) by PCR and then the two genes were cloned into expression vector.

美洲商陆抗病毒蛋白是一类能很好的抑制植物病毒的蛋白，很早就有报道低剂量的商陆叶片粗粉末就能明显的抑制TMV在菜豆叶片上局部枯斑的形成，PAP缓释剂也己用于病毒病的防治工作。

Pokeweed antiviral protein belongs to Ribosome-inactive Proteins. PAP has potent antiviral activity against many plant virus. pT1 was used as the template to amplify the deleted mutant PAP gene (N-deleted and N-,C-deIeted) by PCR and then the two genes were cloned into expression vector. Different IPTG-inducible expression vector containing different deleted mutant PAP gene was constructed and transferred into E.coli strain BL21 (DE3)-plysS.

美洲商陆抗病毒蛋白是一类能很好的抑制植物病毒的蛋白，很早就有报道低剂量的商陆叶片粗粉末就能明显的抑制TMV在菜豆叶片上局部枯斑的形成，PAP缓释剂也已用于病毒病的防治工作，但美洲商陆抗病毒蛋白属于核糖体失活蛋白，它对生长的细胞会有一定的毒性。

Pokeweed antiviral protein, a 29-kD protein isolated from Phytolacca americana, inhibits translation by catalytically removing a specific adenine residue from the large rRNA of the 60s subunit of eukaryotic ribosomes. In addition to its ribosome-inactivating ability, PAP has potent antiviral activity against many plants and animal viruses.

美洲商陆抗病毒蛋白(pokeweed antiviral proteins，PAPs)，是在美洲商陆的不同组织或不同生长阶段生成的一类具酶功能的单链核糖体失活蛋白(ribosome-inactivating proteins，RIP)，是天然的广谱抗病毒生物活性物资，能抗多种植物和动物病毒。

Transgenic tobacco plants were obtained through screening with kanamycin. The transgenic tobacco plants could delay TMV infection for about 25 days compared with non-transgenic tobacco plants. Pokeweed antiviral protein Ⅱ is expressed with high level in summer leaves. The expression of PAPⅡ is regulated by season. The total RNA was extracted from pokeweed (Phytolacca americana L.) leaves in summer using the method of TRIzol and used as template to amplify the PAPⅡ gene by RT-PCR and then the gene was cloned into E. coli expression vector and secreted expression pPIC9K vector. The two vectors with PAPⅡ gene were then transferred into E. coli strain BL21 (DE3)-plysS and Pachia pastoris GS115 strain respectively. The specific protein was produced induced by IPTG and methanol.

由于美洲商陆抗病毒蛋白Ⅱ是一个受季节调控表达的蛋白，本实验以美洲商陆夏季叶片为材料，通过对其总RNA的提取、反转录、并用PAPⅡ的特异引物进行PCR扩增，对PAPⅡ进行了基因克隆、序列分析，结果扩增出来的PAPⅡ基因与已经报道的序列同源性是99.9％，然后将该基因构建到原核、真核表达载体上，分别转化了大肠杆菌和毕赤酵母并对它们进行了诱导表达，在两个表达系统中均获得了有活性的PAPⅡ表达蛋白，体外生物测定表明表达的蛋白均具有抑制病毒的活性，PAPⅡ基因在酵母中还没有表达的报道，这为获得具活性PAPⅡ蛋白提供了一种简便可行的方法。

The transgenic tobacco plants could delay TMV infection for about 25 days compared with non-transgenic tobacco plants.Pokeweed antiviral protein II is expressed with high level in summer leaves. The expression of PAPII is regulated by season. The total RNA was extracted from pokeweed (Phytolacca americana L.) leaves in summer using the method of TRIzol and used as template to amplify the PAPII gene by RT-PCR and then the gene was cloned into E.coli expression vector and secreted expression pPIC9K vector. The two vectors with PAPII gene were then transferred into E.coli strain BL21 (DE3)-plysS and Pachia pastor is GS115 strain respectively. The specific protein was produced induced by IPTG and methanol.

由于美洲商陆抗病毒蛋白Ⅱ是一个受季节调控表达的蛋白，本实验以美洲商陆夏季叶片为材料，通过对其总RNA的提取、反转录、并用PAPⅡ的特异引物进行PCR扩增，对PAPⅡ进行了基因克隆、序列分析，结果扩增出来的PAPⅡ基因与已经报道的序列同源性是99.9％，然后将该基因构建到原核、真核表达载体上，分别转化了大肠杆菌和毕赤酵母并对它们进行了诱导表达，在两个表达系统中均获得了有活性的PAPⅡ表达蛋白，体外生物测定表明表达的蛋白均具有抑制病毒的活性，PAPⅡ基因在酵母中还没有表达的报道，这为获得具活性PAPⅡ蛋白提供了一种简便可行的方法。

The nucleotide sequences of the PAP gene and PAPⅡ gene in the root of Phytolacca americana were determined by reverse transcription polymerase chain reaction and PCR. The gene comprises 942bp and 933bp respectively, encoding 313 amino acids and 310 amino acids. When PAP gene and PAPⅡ gene sequences in the root were compared with those in the spring leaves and summer ones, the highest similarity 100% and 99. 8% were observed respectively. They both had one open reading frame . Analysis of the two sequences showed, in fact, they belonged to one species.

以美洲商陆根为材料，提取其总RNA、反转录，用一对PAP基因的特异性引物和一对PAPⅡ的特异性引物，分别对反转录产物进行PCR扩增，将扩增的目的片段进行序列分析，结果扩增出来的PAP基因与已经报道的序列完全相同，扩增出来的PAPⅡ基因与已经报道的序列同源性也在99.8％，而且它们都各自编码一个开放阅读框架，可以说美洲商陆根中肯定存在PAP和PAPⅡ基因，并已经转录。

The nucleotide sequences of the PAP gene and PAPII gene in the root of Phytolacca americana were determined by reverse transcription polymerase chain reaction and PCR. The gene comprises 942bp and 933bp respectively, encoding 313 amino acids and 310 amino acids. When PAP gene and PAPII gene sequences in the root were compared with those in the spring leaves and summer ones, the highest similarity 100% and 99.8% were observed respectively. They both had one open reading frame. Analysis of the two sequences showed, in fact, they belonged to one species.

以美洲商陆根为材料，提取其总RNA、反转录，用一对PAP基因的特异性引物和一对PAPⅡ的特异性引物，分别对反转录产物进行PCR扩增，将扩增的目的片段进行序列分析，结果扩增出来的PAP基因与已经报道的序列完全相同，扩增出来的PAPⅡ基因与已经报道的序列同源性也在99.8％，而且它们都各自编码一个开放阅读框架，可以说美洲商陆根中肯定存在PAP和PAPⅡ基因，并已经转录。

Pokeweed antiviral protein, a 29-kD protein isolated from Phytolacca americana, inhibits translation by catalytically removing a specific adenine residue from the large rRNA of the 60s subunit of eukaryotic ribosomes. In addition to its ribosome-inactivating ability, PAP has potent antiviral activity against many plants and animal viruses.

美洲商陆抗病毒蛋白(pokeweed antiviral proteins，PAPs)，是在美洲商陆的不同组织或不同生长阶段生成的一类具功能的单链核糖体失活蛋白(ribosome-inactivating proteins，RIP)，是天然的广谱抗病毒生物活性物资，能抗多种植物和动物病毒。

In the negative and interrogative forms, of course, this is identical to the non-emphatic forms.

。但是，在否定句或疑问句里，这种带有"do"的方法表达的效果却没有什么强调的意思。

Go down on one's knees;kneel down

屈膝跪下。。。下跪祈祷