Javaran Vahid Jalali, Shafeinia Alireza, Javaran Mokhtar Jalali, Gojani Esmaeil Ghasemi, Mirzaee Malihe
Department of Biotechnology, Ramin University of Agriculture and Natural Resources, Ahwaz, 63417-711, Iran.
Department of Biotechnology, Tarbiat Modares University, Tehran, 14115-336, Iran.
Biotechnol Lett. 2017 Apr;39(4):607-612. doi: 10.1007/s10529-017-2285-6. Epub 2017 Jan 16.
To use a transient expression system to express a truncated human tissue plasminogen activator (K2S) gene in cucurbit plants.
The recombinant tissue plasminogen activator protein (K2S form) was expressed in active form in cucurbit plants. Its molecular weight was 43 kDa. The plant-derived rt-PA was determined using goat anti-rabbit antibody by western blotting. Among the infected lines, the highest expression of rt-PA was 62 ng/100 mg per leaf tissue as measured by ELISA. The enzymatic activity of the plant-derived rt-PA was 0.8 IU/ml.
The K25 form of rt-PA was expressed for the first time using the viral expression system. Plant-derived rt-PA showed similar potency to commercially-available PA.
利用瞬时表达系统在葫芦科植物中表达截短的人组织纤溶酶原激活剂（K2S）基因。
重组组织纤溶酶原激活剂蛋白（K2S形式）在葫芦科植物中以活性形式表达。其分子量为43 kDa。通过蛋白质免疫印迹法使用山羊抗兔抗体对植物源重组人组织型纤溶酶原激活剂进行测定。在受感染的株系中，通过酶联免疫吸附测定法测得，每100 mg叶片组织中重组人组织型纤溶酶原激活剂的最高表达量为62 ng。植物源重组人组织型纤溶酶原激活剂的酶活性为0.8 IU/ml。
首次利用病毒表达系统表达了K25形式的重组人组织型纤溶酶原激活剂。植物源重组人组织型纤溶酶原激活剂显示出与市售纤溶酶原激活剂相似的效力。
