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A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells
Published on: July 31, 2012
Molecular cloning and expression of human alpha2,8-sialyltransferase (hST8Sia V)
1Molecular Glycobiology Research Unit, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Yusung, Taejon, South Korea.
Abstract:
The cDNA encoding human alpha2,8-sialyltransferase (hST8Sia V) which exhibits activity toward gangliosides, GM1b, GD1a, GT1b, and GD3, was isolated by screening of human brain cDNA library with a DNA probe generated from the cDNA sequence of mouse ST8Sia V (mST8Sia V) and by 5'-RACE of mRNA from human brain tissue. Comparative analysis of this cDNA with mST8Sia V showed that each sequence of the predicted coding region contains 84% identity in both nucleotide and amino acid. Northern analysis of this cDNA indicated that, in contrast to mST8Sia V, two different sizes of transcripts corresponding to 11 and 2.5 kb were expressed in both human fetal and adult brain, while the transcript of 2.5 kb was detected only in adult heart and skeletal muscle. The enzyme expressed in COS cells showed a substrate specificity very similar to that of mST8Sia V.
