In this study,cowpea trypsin inhibitor (CpTI) gene, an insecticidal gene,was introduced into two Populus tomentosa clones by gene transformation mediated by Agrobacterium tumefaciens. The transformed regeneration shoo...In this study,cowpea trypsin inhibitor (CpTI) gene, an insecticidal gene,was introduced into two Populus tomentosa clones by gene transformation mediated by Agrobacterium tumefaciens. The transformed regeneration shoots were obtained directly by leaf discs.In order to established selection condition,leaf discs were cultured in the medium with increasing concentration kanamycin.Kanamycin resistant(km r)plantlets were obtained by 3～4 cycles screening in selective condition.Then transformed shoots were rooted in the medium containing kanamycin 50?mg/L and transferred to greenhouse.The presence of CpTI gene in transgenic plants were confirmed by PCR and PCR\|Southern blot.展开更多
Targeting of the synthesized polypeptide in the cells is an important research field in modern cell biology. Cowpea trypsin inhibitor (cpti) gene has been modified and a fusion protein gene (sck) was produced by fusin...Targeting of the synthesized polypeptide in the cells is an important research field in modern cell biology. Cowpea trypsin inhibitor (cpti) gene has been modified and a fusion protein gene (sck) was produced by fusing a signal peptide sequence at cpti 5' end and an endoplasm reticulum (ER) retention signal peptide at cpti3' end respectively. The signal peptide can direct the newly synthesized polypeptide into ER, while ER retention signal can make the protein retained in the ER and its derivative protein body. ELISA test indicated that the accumulation level of foreign CpTI protein in sck transgenic tobacco (Nicotiana tabacum L.) was two times higher than cpti transgenic tobaccos and some individuals were four times higher. At the same time, sck transgenic tobacco has a high resistance to Lepidoptera pest due to the increased accumulation level of foreign CpTI protein. The strategy of foreign protein targeting can be used to increase the accumulation level of foreign protein in transgenic plants and can be widely applied to other related research field in plant genetic engineering.展开更多
文摘In this study,cowpea trypsin inhibitor (CpTI) gene, an insecticidal gene,was introduced into two Populus tomentosa clones by gene transformation mediated by Agrobacterium tumefaciens. The transformed regeneration shoots were obtained directly by leaf discs.In order to established selection condition,leaf discs were cultured in the medium with increasing concentration kanamycin.Kanamycin resistant(km r)plantlets were obtained by 3～4 cycles screening in selective condition.Then transformed shoots were rooted in the medium containing kanamycin 50?mg/L and transferred to greenhouse.The presence of CpTI gene in transgenic plants were confirmed by PCR and PCR\|Southern blot.
文摘Targeting of the synthesized polypeptide in the cells is an important research field in modern cell biology. Cowpea trypsin inhibitor (cpti) gene has been modified and a fusion protein gene (sck) was produced by fusing a signal peptide sequence at cpti 5' end and an endoplasm reticulum (ER) retention signal peptide at cpti3' end respectively. The signal peptide can direct the newly synthesized polypeptide into ER, while ER retention signal can make the protein retained in the ER and its derivative protein body. ELISA test indicated that the accumulation level of foreign CpTI protein in sck transgenic tobacco (Nicotiana tabacum L.) was two times higher than cpti transgenic tobaccos and some individuals were four times higher. At the same time, sck transgenic tobacco has a high resistance to Lepidoptera pest due to the increased accumulation level of foreign CpTI protein. The strategy of foreign protein targeting can be used to increase the accumulation level of foreign protein in transgenic plants and can be widely applied to other related research field in plant genetic engineering.
