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Molecular cloning of the phenylalanine ammonia lyase gene from Rhodosporidium toruloides in Escherichia coli K-12

Molecular cloning of the phenylalanine ammonia lyase gene from Rhodosporidium toruloides in Escherichia coli K-12
Molecular cloning of the phenylalanine ammonia lyase gene from Rhodosporidium toruloides in Escherichia coli K-12
A genomic library of Rhodosporidium toruloides DNA was constructed in bacteriophage lambda 1059. Recombinant phage containing phenylalanine ammonia lyase (PAL) gene sequences were identified by using 32P-labeled cDNA to partially purified PAL mRNA. The PAL gene was subcloned on an 8.5-kilobase PstI DNA restriction fragment into pUC8 to generate the recombinant plasmid pHG2. A restriction map of the PAL gene, together with its flanking regions, was constructed. Northern hybridization analysis of R. toruloides RNA with a restriction fragment encoding part of the PAL gene indicates that PAL mRNA is 2.5 kilobases in length. A single-stranded DNA hybridization probe was constructed and used to quantitate PAL mRNA levels in R. toruloides grown under different physiological conditions. PAL mRNA levels paralleled changes in functional PAL mRNA and antigen. These data are consistent with control of PAL expression being at the level of transcription.
0021-9193
314-320
Gilbert, Harry J.
462d2685-21e2-4aac-a6b4-a051923ee42a
Clarke, Ian N.
ff6c9324-3547-4039-bb2c-10c0b3327a8b
Gibson, Raymond K.
285c7b76-75e1-4acb-b5f6-d3bb902c646c
Stephenson, John R.
d4e208fd-763c-418a-b4a0-f2e2c90862cc
Tully, Michael
e02c4036-b0ee-4e5a-976d-aee0b38b81bd
Gilbert, Harry J.
462d2685-21e2-4aac-a6b4-a051923ee42a
Clarke, Ian N.
ff6c9324-3547-4039-bb2c-10c0b3327a8b
Gibson, Raymond K.
285c7b76-75e1-4acb-b5f6-d3bb902c646c
Stephenson, John R.
d4e208fd-763c-418a-b4a0-f2e2c90862cc
Tully, Michael
e02c4036-b0ee-4e5a-976d-aee0b38b81bd

Gilbert, Harry J., Clarke, Ian N., Gibson, Raymond K., Stephenson, John R. and Tully, Michael (1985) Molecular cloning of the phenylalanine ammonia lyase gene from Rhodosporidium toruloides in Escherichia coli K-12. Journal of Bacteriology, 161 (1), 314-320. (PMID:7041889)

Record type: Article

Abstract

A genomic library of Rhodosporidium toruloides DNA was constructed in bacteriophage lambda 1059. Recombinant phage containing phenylalanine ammonia lyase (PAL) gene sequences were identified by using 32P-labeled cDNA to partially purified PAL mRNA. The PAL gene was subcloned on an 8.5-kilobase PstI DNA restriction fragment into pUC8 to generate the recombinant plasmid pHG2. A restriction map of the PAL gene, together with its flanking regions, was constructed. Northern hybridization analysis of R. toruloides RNA with a restriction fragment encoding part of the PAL gene indicates that PAL mRNA is 2.5 kilobases in length. A single-stranded DNA hybridization probe was constructed and used to quantitate PAL mRNA levels in R. toruloides grown under different physiological conditions. PAL mRNA levels paralleled changes in functional PAL mRNA and antigen. These data are consistent with control of PAL expression being at the level of transcription.

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Published date: January 1985
Organisations: Faculty of Medicine

Identifiers

Local EPrints ID: 352621
URI: https://eprints.soton.ac.uk/id/eprint/352621
ISSN: 0021-9193
PURE UUID: 9bb551f9-514b-4578-a498-96e2fe67c062
ORCID for Ian N. Clarke: ORCID iD orcid.org/0000-0002-4938-1620

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Date deposited: 30 May 2013 11:27
Last modified: 06 Jun 2018 13:19

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Contributors

Author: Harry J. Gilbert
Author: Ian N. Clarke ORCID iD
Author: Raymond K. Gibson
Author: John R. Stephenson
Author: Michael Tully

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