Mutation and gene transfer of neutral amino acid transport System L genes in mammalian cells
The authors are attempting to clone the genes coding for amino acid transport System L. Chinese hamster ovary (CHO) cell mutants that are temperature sensitive in their leucyl-tRNA synthetase show temperature-dependent regulation of System L. Temperature resistant mutants isolated from these cells have constitutively derepressed System L activity. Somatic cell fusion studies using these mutants have suggested that a trans-acting element controls regulation of System L. Mutants with reduced transport activity were isolated by a TH-suicide selection. The growth of these mutant cells is limited by the transport defect. CHO mutants were transformed with a human cosmid library, followed by selection at high temperatures and low leucine concentrations. Some transformants have increased levels of System L activity, suggesting that human genes coding for leucine transport have been incorporated into the CHO genome. Human sequences were rescued by a lambda in vitro packaging system. These sequences hybridize to vector and total human DNA. Experiments are being done to confirm that these sequences indeed code for transport System L. They are also attempting to label membrane components of amino acid transporters by group-specific modifying reagents.
- Research Organization:
- Univ. of Michigan, Ann Arbor
- OSTI ID:
- 6155661
- Report Number(s):
- CONF-870644-; TRN: 87-028772
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CHO CELLS
GENE REGULATION
SENSITIVITY
GENES
CLONING
LEUCINE
MEMBRANE TRANSPORT
AMINO ACID SEQUENCE
CELL MEMBRANES
DNA
MUTANTS
MUTATIONS
SOMATIC CELLS
TEMPERATURE DEPENDENCE
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AMINO ACIDS
ANIMAL CELLS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MEMBRANES
MOLECULAR STRUCTURE
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
550401* - Genetics- Tracer Techniques