Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 3 1 2011 05 22 Unexpected Transcription of Human t-PA cDNA in transgenic tobacco plants Unexpected Transcription of Human t-PA cDNA in transgenic tobacco plants 31 41 459 10.22103/jab.2012.459 FA Mokhtar Jalali 0000-0002-4668-0764 Heydar Seifi Nabiabad Mohammad Mehdi Yaghoobi Saman Hossein Khani Journal Article 2011 03 02 Recombinant tissue-type plasminogen activator (rt-PA) is used as a fibrin-specific thrombolytic agent for the treatment of various thromboembolic diseases. In this study, we tried to show the expression of transferred cDNA of human t-PA gene in transgenic tobacco plants produced previously by this group. RT-PCR with specific primers for t-PA, were used to assay the transgenic plants. RT-PCR products were observed on agarose gel electrophoresis after ethidium bromide staining. The results showed that the t-PA gene is stably expressed in second generation transgenic plant at the level of mRNA, while no expression was seen in non transgenic plants. These results also indicate an unfamiliar 650 bp transcript from t-PA transgene in plants. The results of homology search by CDD at NCBI database demonstrate that in addition to complete cDNA, an incomplete transcript so observed. This truncated transcript is a part of cDNA of t-PA that has lost the kringles domain while tyrosine-like serine protease is remained. This may results from alternative RNA splicing. The serine protease domain can carry out the role of t-PA alone. So many scientists are trying to clone this fragment only as a drug. Interestingly the plant produces this fragment naturally. This result may be because of alternative splicing of RNA. Recombinant tissue-type plasminogen activator (rt-PA) is used as a fibrin-specific thrombolytic agent for the treatment of various thromboembolic diseases. In this study, we tried to show the expression of transferred cDNA of human t-PA gene in transgenic tobacco plants produced previously by this group. RT-PCR with specific primers for t-PA, were used to assay the transgenic plants. RT-PCR products were observed on agarose gel electrophoresis after ethidium bromide staining. The results showed that the t-PA gene is stably expressed in second generation transgenic plant at the level of mRNA, while no expression was seen in non transgenic plants. These results also indicate an unfamiliar 650 bp transcript from t-PA transgene in plants. The results of homology search by CDD at NCBI database demonstrate that in addition to complete cDNA, an incomplete transcript so observed. This truncated transcript is a part of cDNA of t-PA that has lost the kringles domain while tyrosine-like serine protease is remained. This may results from alternative RNA splicing. The serine protease domain can carry out the role of t-PA alone. So many scientists are trying to clone this fragment only as a drug. Interestingly the plant produces this fragment naturally. This result may be because of alternative splicing of RNA. https://jab.uk.ac.ir/article_459_7fe1f8abaad094e0b5cb1b01d712f708.pdf