1.Expression Of Human Growth Hormone In Silkworm Cell 2.Inducible Activity Of Antimicrobial Peptide Promoter In Insect Cell

Human growth hormone( hGH) is a kind of important polypeptide produced by the anterior portion of the pituitary gland, which could facilitate the growth of the bone and promote the metabolism of the organism. And the maintenance of blood hGH level is vital for the normal growth and. Human body's modulation for the surrounding. The true meaning of the gene engineering to express the hGH is therapy of the dwarf and ectogenesis complementation to promote the blood hGH when the atrophy of old people's pituitary gland begins. Compared to the prokaryotic gene expressing systems and mammalian gene expressing systems, insect gene expressing systems possess the stronger ability of the transcription and post-translation processing, which also has high production. We can anticipate it will be a kind of potent effective ectogenesis eukaryotic gene expressing system. At present, most of the gene expression is often associated with attempts to express cDNA constructs. However, someone has confirmed that a striking improvement both in the number of the transgenic mice that expressed the gene and in the average level of expression when the natural introns were included. In addition, some experiments indicated that the introns had a major effects on transcription rather than mRNA processing or stability. As an insect expressing system, silkworm has long been thought highly of. However, because the relation between insects and mammals is in evolution is so distant, we do not have the exact idea if the introns of the mammalian gene can be spliced correctly in insect cells and what influence the intron imposed upon the gene expressed in the insect cells. Therefore, we hope to construct a effective eukaryotic gene expressing vector harboring a genomic DNA, including introns, and develop a gene expressing system could correctly splice the mRNA. We generated a recombinant eukaryotic gene expressing vector harboring a full-length hGH gene, 2.4 kb genomic DNA with four introns and the signal peptide sequence cloned to the eukaryotic gene expressing vector pcDNA3.0. Then we transfected the plasmid to the BmN cells through lipofectin. First, we extract the cell's total RNA and amplify the cDNA through RT PCR , which showed the presence of RNA corresponding to the authetic hGH mRNA. During the different time point, we retrieve the cell supernatant and through the electrophoretic analysis and immunoblot of the recombinant protein indicated that the cell which transfected the recombinant eukaryotic gene expressing vector could synthesis and excrete the peptide effectively. Our data indicate that in the insect cells, we can express the human full-length gene carrying the introns and signal peptide.