1993 Fiscal Year Final Research Report Summary

The analysis of monocyte chemoattractant protein-1(MCP-1) receptor

Research Project

Project/Area Number	04670394
Research Category	Grant-in-Aid for General Scientific Research (C)
Allocation Type	Single-year Grants
Research Field	内科学一般
Research Institution	Yokohama City University
Principal Investigator	TANI Kenji 横浜市立大学, 附属浦舟病院第一内科, 助教授 (20094310)
Project Period (FY)	1992 – 1993
Keywords	MCP-1 / baculovirus system
Research Abstract	Human monocyte chemoattractant protein-1 (hMCP-1) was produced using baculovirus system. The hMCP-1 cDNA was inserted into the genomic DNA of Autographa californica nuclear polyhedrosis virus (AcNPV) using a transfer vector, pJVP10Z, Spodoptera frugiperda insect cells, which were infected with this recombinant virus, secreted recombinant hMCP-1 (re-hMCP-1) at the level of 10-20mug/ml of culture medium. This product was shown to chemoattract monocytes. Three distinet bands of 11, 11.5 and 12 kDa were revealed by immunoblotting analysis, and this heterogeneity was assigned to differences in carbohydrate processing. N-terminal amino acid sequence analysis of the purified product revealed identity with hMCP-1. Thus, in this system, re-hMCP-1 was produced in large quantities and modified in a manner similar to native hMCP-1.