Ueda A, Kawamoto S, Igarashi T, Ishigatsubo Y, Tani K, Okubo T, Okuda K
Department of Internal Medicine I, Yokohama City University School of Medicine, Japan.
Gene. 1994 Mar 25;140(2):267-72. doi: 10.1016/0378-1119(94)90556-8.
Human monocyte chemoattractant protein-1 (hMCP-1) was produced using a 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 10-20 micrograms/ml of culture medium. This product was shown to chemoattract monocytes. Three distinct 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.
人单核细胞趋化蛋白-1（hMCP-1）是利用杆状病毒系统生产的。使用转移载体pJVP10Z将hMCP-1 cDNA插入苜蓿银纹夜蛾核型多角体病毒（AcNPV）的基因组DNA中。用这种重组病毒感染的草地贪夜蛾昆虫细胞，在培养基中以10-20微克/毫升的水平分泌重组hMCP-1（re-hMCP-1）。该产物显示出对单核细胞的趋化作用。免疫印迹分析显示出11、11.5和12 kDa的三条不同条带，这种异质性归因于碳水化合物加工的差异。纯化产物的N端氨基酸序列分析显示与hMCP-1相同。因此，在该系统中，大量生产了re-hMCP-1，并以与天然hMCP-1相似的方式进行了修饰。
