By R. E. Spier
This booklet provides a state of the art document at the box of animal mobilephone know-how, a resource of reference for all these fascinated with the construction and use of animal phone items
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Extra info for Animal Cell Technology. Developments, Processes and Products
After fos+myb+myc IMP * selection, the IL-6 produc fos+ras+myc ! tivities of cells per each well myb+ras+myc were determined. mt-" r w w myb+ras+myc \ r* 57 The expression level of mRNA was examined by Northern blotting. No sig nal of myb mRNA was detectable for SFME cells. A^/>-transfected SFME cells were estimated to express about 6 copies of mRNA per single cells. Appearance of highly IL-6 productive clones by transfection of BHK-21 cells expressing the IL-6 gene with oncogenes. BHK-21 cells are widely used for production of bioactive proteins.
Human IL-6 cDNA were purchased from Toyobo Co. The pSRcr -IL6 was con structed by recombinating the human IL-6 cDNA containing leader sequence into EcoRI site of pcDL-SRa 296 vector. Human or fos, avian v-myb, human c-Ha-ras, human c-myc genomic oncogenes were obtained from Japanese Can cer Research Resources Bank (JCRB). These oncogenes were inserted in the multicloning site of pRC/CMV vector, as described previously4. Transfection procedure SFME cells were transfected with high efficiency calcium phosphate coprecipitation method5.
However, with the 3 day subcultures, it appears that the growth of the cells in spinners was less than that of cells in fermenters. This difference was almost certainly the consequence of poor culture conditions in the spinners, such as a reduced pH or DOT leading to an inhibition of cell growth during the third day of culture. No trend with time was observed in terms of change in maximum cell numbers for the spinner or fermenter cultures. However, a period of adaptation of the cells to fermenter culture was seen, during which time the cells ceased to grow in a clumped manner and grew as a single cell suspension (see below).
Animal Cell Technology. Developments, Processes and Products by R. E. Spier