We stained cell lines with lectins that have been used to recognize the following three different epitopes: (i) WGA recognizes all sialic acids, (ii) MAA recognizes α2,3 sialic acid, and (iii) SNA recognizes α2,6 sialic acid. In this report, we demonstrate that AAV1 and AAV6 use both α2,3 and α2,6 N-linked sialic acids for binding and infection. To analyze if the sialylated receptors of AAV1 and AAV6 are glycoproteins or glycolipids, we treated Cos-7 and Pro-5 cells with 200 μg/ml proteinase K. Cell viability was assessed immediately after proteinase K treatment, and no cytotoxic effect was observed. Sialic acids are abundant on the surfaces of muscle cells, which may partly explain the high transduction efficiency of AAV1 and AAV6 vectors for this tissue. To determine if N-linked sialic acid is necessary for efficient AAV1 and AAV6 transduction, we tested AAV1 and AAV6 transduction on another Pro-5-derived cell line, Lec-1 (40), that is deficient in N-linked glycans but is not deficient in O-linked glycans (Fig. (Fig.7C).7C). Resialylation by α2,3(N)-sialyltransferase and α2,6(N)-sialyltransferase increased AAV6 transduction by 11-fold and 6-fold, respectively, but only 2.5-fold for AAV1. Therefore, Pro-5 cells and HepG2 cells display α2,3 sialic acid or α2,6 sialic acid on their surfaces, respectively, while Cos-7 cells display α2,3 sialic acid and a relatively small amount of α2,6 sialic acid.
The following sialyltransferases were used to add specific sialic acids to the surfaces of Lec-2 cells: α2,3(O)-sialyltransferase, α2,3(N)-sialyltransferase, and α2,6(N)-sialyltransferase. The page "Sialic acid manufacturer" does not exist. ’ says the swine-bird-human flu strain, reported to be found first in Mexico in late-March 2009, could have only come from Dr James S. Robertson and his colleagues in association with the US Centre for Disease Control and vaccine manufacturer Novavax, Inc, which was ready to profit from the release he says. Sialic Acid or N-Acetylneuraminic Acid Sialic Acid or N-Acetylneuraminic Acid factory, Supplier, Manufacturer. All glycans recognized contain a common motif: sialic acid linked to N-acetyl-lactosamine. A total of 264 glycans were screened for binding to the capsids as described in Materials and Methods. For example, methods of transforming Bacillus species and promoters that can be used to express proteins are taught in U.S. Such promoters can be obtained from genes that have been cloned from the species, or heterologous promoters can be used. Promoters for use in E. coli include the T7, trp, or lambda promoters.
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