Abstract
Twelve complementary DNA clones for human lysosomal α- galactosidase A were isolated from an Okayama-Berg library constructed from SV40-transformed human fibroblasts. The identity of these clones was confirmed by complete colinearity of the nucleotide-deduced amino acid sequence with that determined by direct chemical sequencing of human placental α-galactosidase A. Hybridization of the α-galactosidase A cDNA to genomic DNA from individuals with varying numbers of X chromosomes as well as from interspecies somatic-cell hybrids showed only a single locus in the genoma at Xq 13.1 - Xq 22. One cDNA clone (pcD-AG210) contained the complete coding sequence for both the signal peptide and mature α-galactosidase A. The signal peptide of 31 amino acids contains the expected hydrophobic domains consisting of Leu-Gly-Cys-Ala-Leu-Ala-Leu and Phe-Leu-Ala-Leu-Val and has Ala at the signal peptidase cleavage site. Twelve out of 15 G residues flanking the 5' end of the cDNA in pcD-AG210 were removed and the truncated fragment was ligated into the original vector. This construct, pcD-AG502, encoded enzymatically active human α-galactosidase A in monkey COS cells.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 275-280 |
| Number of pages | 6 |
| Journal | European Journal of Biochemistry |
| Volume | 165 |
| Issue number | 2 |
| DOIs | |
| State | Published - 1987 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biochemistry
Cite this
Signal sequence and DNA-mediated expression of human lysosomal α-galactosidase A. / Tsuji, S.; Martin, B. M.; Kaslow, D. C.; Migeon, B. R.; Choudary, Prabhakara V; Stubbleflied, B. K.; Mayor, J. A.; Murray, G. J.; Barranger, J. A.; Ginns, E. I.
In: European Journal of Biochemistry, Vol. 165, No. 2, 1987, p. 275-280.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Signal sequence and DNA-mediated expression of human lysosomal α-galactosidase A
AU - Tsuji, S.
AU - Martin, B. M.
AU - Kaslow, D. C.
AU - Migeon, B. R.
AU - Choudary, Prabhakara V
AU - Stubbleflied, B. K.
AU - Mayor, J. A.
AU - Murray, G. J.
AU - Barranger, J. A.
AU - Ginns, E. I.
PY - 1987
Y1 - 1987
N2 - Twelve complementary DNA clones for human lysosomal α- galactosidase A were isolated from an Okayama-Berg library constructed from SV40-transformed human fibroblasts. The identity of these clones was confirmed by complete colinearity of the nucleotide-deduced amino acid sequence with that determined by direct chemical sequencing of human placental α-galactosidase A. Hybridization of the α-galactosidase A cDNA to genomic DNA from individuals with varying numbers of X chromosomes as well as from interspecies somatic-cell hybrids showed only a single locus in the genoma at Xq 13.1 - Xq 22. One cDNA clone (pcD-AG210) contained the complete coding sequence for both the signal peptide and mature α-galactosidase A. The signal peptide of 31 amino acids contains the expected hydrophobic domains consisting of Leu-Gly-Cys-Ala-Leu-Ala-Leu and Phe-Leu-Ala-Leu-Val and has Ala at the signal peptidase cleavage site. Twelve out of 15 G residues flanking the 5' end of the cDNA in pcD-AG210 were removed and the truncated fragment was ligated into the original vector. This construct, pcD-AG502, encoded enzymatically active human α-galactosidase A in monkey COS cells.
AB - Twelve complementary DNA clones for human lysosomal α- galactosidase A were isolated from an Okayama-Berg library constructed from SV40-transformed human fibroblasts. The identity of these clones was confirmed by complete colinearity of the nucleotide-deduced amino acid sequence with that determined by direct chemical sequencing of human placental α-galactosidase A. Hybridization of the α-galactosidase A cDNA to genomic DNA from individuals with varying numbers of X chromosomes as well as from interspecies somatic-cell hybrids showed only a single locus in the genoma at Xq 13.1 - Xq 22. One cDNA clone (pcD-AG210) contained the complete coding sequence for both the signal peptide and mature α-galactosidase A. The signal peptide of 31 amino acids contains the expected hydrophobic domains consisting of Leu-Gly-Cys-Ala-Leu-Ala-Leu and Phe-Leu-Ala-Leu-Val and has Ala at the signal peptidase cleavage site. Twelve out of 15 G residues flanking the 5' end of the cDNA in pcD-AG210 were removed and the truncated fragment was ligated into the original vector. This construct, pcD-AG502, encoded enzymatically active human α-galactosidase A in monkey COS cells.
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UR - http://www.scopus.com/inward/citedby.url?scp=0023273544&partnerID=8YFLogxK
U2 - 10.1111/j.1432-1033.1987.tb11438.x
DO - 10.1111/j.1432-1033.1987.tb11438.x
M3 - Article
C2 - 3036505
AN - SCOPUS:0023273544
VL - 165
SP - 275
EP - 280
JO - FEBS Journal
JF - FEBS Journal
SN - 1742-464X
IS - 2
ER -