Expression And Purification Of TAT/CT-1 And TAT/EGFP Fusion Proteins

Nowadays, spinal cord injury grasp great attention in our society, it is very important to intensify the cure of SCI. In all of the measures, neurotrophic factors (NTF) promote the survival of neuron after SCI, and up-regulate related gene of regeneration.Cardiotrophin-1(CT-1) was isolated as a new cytokine by Pennica in 1995, it was constructed by 201 amino acid, and can stimulate the growth of neonate rat' cardiac muscle in vitro, and belong to the interleukin (IL)-6-related cytokine family. Because of the homology to CNTF, CT-1 grasp more and more people's attention.CT-1 can sustain the survival of motor neuron (including spinal and brain motor neuron), dihydroxyphenyl ethylamine neuron and sympathetic neuron. Some people have already reported the gene therapy of rat progressive motor neuron disease and the stiffen of spinal cord's lateral bundle by CT-1, and get good effects. Zhang Zhengfeng has cloned human CT-1 gene, and expressed, purified recombinant CT-1 protein, construct recombinant adenovirus vector using human CT-1, and using it in the research of nerve injury. In the aspect of peripheral nerve injury, recombinant CT-1 protein can promote the survival of motor neurone in cornu anterius medullae spinalis.In the meanwhile, because of the neurotrophic factors' inability to transit blood-brain barrier and it's short demiperiod, So providing high-performance neurotrophic factors to the area of SCI and protecting regenerating neuron is the popular project, and the measure of long-term,repeated and convenient dosage is the difficult problem of the research of SCI.Trans-activator (Tat) protein coded by human immunodeficiency virus, (HIV-1) contain a protein transduction related peptide fragment, named protein transduction domain. The protein transduction mediated by Tat, isn't dependent acceptor and Transport protein, and do not need power, and can proceed at 4°C. Now it is believed that this process is related to the basic amino acid (Arg and lysine) in protein transduction domain. These amino acids contain strong positive charge, and it may interact with the cellular membrane...