CLONING AND CHARACTERIZATION OF A GOLGI-ASSOCIATED GTP-BINDING PROTEIN HOMOLOG FROM LEISHMANIA-MAJOR
- Author(s)
- Cappai, R; Osborn, AH; Gleeson, PA; Handman, E;
- Details
- Publication Year 1993-11,Volume 62,Issue #1,Page 73-82
- Journal Title
- MOLECULAR AND BIOCHEMICAL PARASITOLOGY
- Publication Type
- Journal Article
- Abstract
- This paper describes the cloning of a Golgi-associated GTP-binding protein homologue from Leishmania major. The gene was isolated using degenerate oligonucleotides to conserved sequences amongst the small GTP-binding proteins in a polymerase chain reaction on genomic DNA of the L. major cloned line V121. The reading frame of one clone showed high similarity to the rab/YPT subfamily of small GTP-binding proteins. A full length copy of the gene was isolated from a lambda gt10 V121 genomic library and sequenced. At the amino acid level the gene showed highest similarity to the human/rat rab1A gene and the mouse/yeast YPT gene and was named LmYPT. The LmYPT gene was present as a single copy gene in both the L. major and L. donovani genomes. Karyotype analysis localized the LmYPT gene to chromosome band 18 in V121, but it was located on a larger chromosome in the different L. major isolate L119. The LmYPT gene was transcribed as a 3.9-kb transcript in both the promastigote and amastigote forms of the parasite. Western blot analysis, using a polyclonal rabbit antiserum raised against an Escherichia coli expressed portion of the molecule, identified a doublet at 20 and 23 kDa in total promastigote protein. Immunoelectron microscopy in combination with peroxidase staining localized the LmYPT molecule to the Leishmania Golgi apparatus.
- Publisher
- ELSEVIER SCIENCE BV
- Keywords
- ADENYLATE-CYCLASE; MOLECULAR-CLONING; ONCOGENE PRODUCT; YPT1 PROTEIN; YEAST YPT1; GENE; FAMILY; PURIFICATION; SUPERFAMILY; KARYOTYPE
- Publisher's Version
- https://doi.org/10.1016/0166-6851(93)90179-2
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 1993-11-01 12:00:00