Porcine α-1,3-galactosyltransferase:: full length cDNA cloning, genomic organization, and analysis of splicing variants

被引:16
作者
Katayama, A
Ogawa, H
Kadomatsu, K
Kurosawa, N
Kobayashi, T
Kaneda, N
Uchimura, K
Yokoyama, I
Muramatsu, T [1 ]
Takagi, H
机构
[1] Nagoya Univ, Sch Med, Dept Biochem, Showa Ku, Nagoya, Aichi 4668990, Japan
[2] Nagoya Univ, Sch Med, Dept Surg 2, Showa Ku, Nagoya, Aichi 4668990, Japan
关键词
alpha 1,3-galactosyltransferase gene; splicing variants; xenotransplantation;
D O I
10.1023/A:1006963809894
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Full length cDNA and genomic DNA of porcine alpha-1,3-galactosyltransferase were isolated, and their structures were analysed. The coding region was encoded by six exons as in the mouse, and the length of each exon was conserved between the two species. The porcine exons were designated Exon 4-9, since in the mouse coding exons started from Exon 4. Introns tended to be longer in the porcine gene; the distance from Exon 4 to the 3'-end of Exon 9 was 24 kb, while this region was 18 kb in the mouse gene. The cDNA structure was extended from the previous data to the 3'-end and to the 5' side of the cDNA. In addition to a cDNA clone with all coding exons, clones lacking parts of these exons were isolated and their structures were determined. One variant lacked Exon 5; the second, Exons 5 and 6; and the third, Exons 5, 6 and 7. The last variant was not found in the mouse, and cDNA transfection of this variant yielded scarcely any enzymatic activity using asialo al-acid glycoprotein as a substrate, and decreased activity using N-acetyllactosamine as a substrate.
引用
收藏
页码:583 / 589
页数:7
相关论文
共 20 条
[1]  
BREATHNACH R, 1981, ANNU REV BIOCHEM, V50, P349, DOI 10.1146/annurev.bi.50.070181.002025
[2]   EFFECTIVE AMPLIFICATION OF LONG TARGETS FROM CLONED INSERTS AND HUMAN GENOMIC DNA [J].
CHENG, S ;
FOCKLER, C ;
BARNES, WM ;
HIGUCHI, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (12) :5695-5699
[3]  
CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2
[4]   RAPID PRODUCTION OF FULL-LENGTH CDNAS FROM RARE TRANSCRIPTS - AMPLIFICATION USING A SINGLE GENE-SPECIFIC OLIGONUCLEOTIDE PRIMER [J].
FROHMAN, MA ;
DUSH, MK ;
MARTIN, GR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (23) :8998-9002
[5]   A UNIQUE NATURAL HUMAN-IGG ANTIBODY WITH ANTI-ALPHA-GALACTOSYL SPECIFICITY [J].
GALILI, U ;
RACHMILEWITZ, EA ;
PELEG, A ;
FLECHNER, I .
JOURNAL OF EXPERIMENTAL MEDICINE, 1984, 160 (05) :1519-1531
[6]   EVOLUTIONARY RELATIONSHIP BETWEEN THE NATURAL ANTI-GAL ANTIBODY AND THE GAL-ALPHA-1-]3GAL EPITOPE IN PRIMATES [J].
GALILI, U ;
CLARK, MR ;
SHOHET, SB ;
BUEHLER, J ;
MACHER, BA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1987, 84 (05) :1369-1373
[7]   DEFINING THE MINIMAL SIZE OF CATALYTICALLY ACTIVE PRIMATE ALPHA-1,3 GALACTOSYLTRANSFERASE - STRUCTURE-FUNCTION STUDIES ON THE RECOMBINANT TRUNCATED ENZYME [J].
HENION, TR ;
MACHER, BA ;
ANARAKI, F ;
GALILI, U .
GLYCOBIOLOGY, 1994, 4 (02) :193-201
[8]  
JOZIASSE DH, 1992, J BIOL CHEM, V267, P5534
[9]  
JOZIASSE DH, 1989, J BIOL CHEM, V264, P14290
[10]   ISOLATION OF A CDNA-ENCODING A MURINE UDPGALACTOSE-BETA-D-GALACTOSYL-1,4-N-ACETYL-D-GLUCOSAMINIDE ALPHA-1,3-GALACTOSYLTRANSFERASE - EXPRESSION CLONING BY GENE-TRANSFER [J].
LARSEN, RD ;
RAJAN, VP ;
RUFF, MM ;
KUKOWSKALATALLO, J ;
CUMMINGS, RD ;
LOWE, JB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (21) :8227-8231