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Response of Escherichia coli Containing Mycobacterial Carotene Genes to UV Radiation.

Houssaini-Iraqui M, Khamlichi N, El Yamani J, Rastogi N - J. Biomed. Biotechnol. (2001)

Bottom Line: The plasmid pC5, which encodes biogenesis of lycopene in Mycobacterium aurum A(+), was partially digested by restriction endonucleases and generated fragments were cloned.After transformation of Escherichia coli (colorless bacteria) with the plasmids so constructed, seven orange clones were detected and found to carry the same recombinant plasmid (pC51).E. coli cells containing this plasmid synthesize neurosporene and lycopene, and were more resistant to ultraviolet irradiation than non pigmented strain.

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ABSTRACT
The plasmid pC5, which encodes biogenesis of lycopene in Mycobacterium aurum A(+), was partially digested by restriction endonucleases and generated fragments were cloned. After transformation of Escherichia coli (colorless bacteria) with the plasmids so constructed, seven orange clones were detected and found to carry the same recombinant plasmid (pC51). E. coli cells containing this plasmid synthesize neurosporene and lycopene, and were more resistant to ultraviolet irradiation than non pigmented strain.

No MeSH data available.


Related in: MedlinePlus

Physical map of pC51 plasmid DNA and its alignment with the pC5 restriction map. Dots represent the plasmid vector pHLD69. Symbol; pr: Z promoter, and the arrow indicates its orientation. Restriction endonuclease sites are abbreviated as follows; B, BamHI; E, EcoR I; K, Kpn I; P, Pst I; S, Sph I. The ORFs A, B, and C were identified on the basis of sequence homologies, and encode for GGPP synthase, phytoene desaturase and phytoene synthase, respectively [6].
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Figure 2: Physical map of pC51 plasmid DNA and its alignment with the pC5 restriction map. Dots represent the plasmid vector pHLD69. Symbol; pr: Z promoter, and the arrow indicates its orientation. Restriction endonuclease sites are abbreviated as follows; B, BamHI; E, EcoR I; K, Kpn I; P, Pst I; S, Sph I. The ORFs A, B, and C were identified on the basis of sequence homologies, and encode for GGPP synthase, phytoene desaturase and phytoene synthase, respectively [6].

Mentions: Clones of E. coli containing plasmid pC5 (Figure 2) did not form any carotene [5, 18]. This plasmid was partially digested by the restriction endonucleases Sau 3A, Sph I, or Pst I. The fragments generated were cloned, respectively, in the pHLD69 vector digested with Bam HI or Sph I or Pst I. The three ligation mixtures were used to transform E. coli (nonpigmented bacterium). After transformation, bacteria were transferred to solid media containing IPTG. All the clones obtained by the cloning with the enzymes Bam HI or Sph I were nonpigmented. Amongst those obtained by the cloning with Pst I, seven orange clones were detected and cloned. Plasmids DNAs were prepared from these orange colonies and subjected to Pst I, Sph I, Bam HI digestion. This experiment (results not shown) showed that all the clones harbored the same recombinant plasmid designated pC51 containing an insert of 4.42 kb (Figure 2). Further, E. coli was transformed by pC51, and all the clones obtained after 60 h of incubation at 37°C were orange-colored. It was therefore concluded that the plasmid pC51 is responsible for the orange pigmentation of the bacteria.


Response of Escherichia coli Containing Mycobacterial Carotene Genes to UV Radiation.

Houssaini-Iraqui M, Khamlichi N, El Yamani J, Rastogi N - J. Biomed. Biotechnol. (2001)

Physical map of pC51 plasmid DNA and its alignment with the pC5 restriction map. Dots represent the plasmid vector pHLD69. Symbol; pr: Z promoter, and the arrow indicates its orientation. Restriction endonuclease sites are abbreviated as follows; B, BamHI; E, EcoR I; K, Kpn I; P, Pst I; S, Sph I. The ORFs A, B, and C were identified on the basis of sequence homologies, and encode for GGPP synthase, phytoene desaturase and phytoene synthase, respectively [6].
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC113779&req=5

Figure 2: Physical map of pC51 plasmid DNA and its alignment with the pC5 restriction map. Dots represent the plasmid vector pHLD69. Symbol; pr: Z promoter, and the arrow indicates its orientation. Restriction endonuclease sites are abbreviated as follows; B, BamHI; E, EcoR I; K, Kpn I; P, Pst I; S, Sph I. The ORFs A, B, and C were identified on the basis of sequence homologies, and encode for GGPP synthase, phytoene desaturase and phytoene synthase, respectively [6].
Mentions: Clones of E. coli containing plasmid pC5 (Figure 2) did not form any carotene [5, 18]. This plasmid was partially digested by the restriction endonucleases Sau 3A, Sph I, or Pst I. The fragments generated were cloned, respectively, in the pHLD69 vector digested with Bam HI or Sph I or Pst I. The three ligation mixtures were used to transform E. coli (nonpigmented bacterium). After transformation, bacteria were transferred to solid media containing IPTG. All the clones obtained by the cloning with the enzymes Bam HI or Sph I were nonpigmented. Amongst those obtained by the cloning with Pst I, seven orange clones were detected and cloned. Plasmids DNAs were prepared from these orange colonies and subjected to Pst I, Sph I, Bam HI digestion. This experiment (results not shown) showed that all the clones harbored the same recombinant plasmid designated pC51 containing an insert of 4.42 kb (Figure 2). Further, E. coli was transformed by pC51, and all the clones obtained after 60 h of incubation at 37°C were orange-colored. It was therefore concluded that the plasmid pC51 is responsible for the orange pigmentation of the bacteria.

Bottom Line: The plasmid pC5, which encodes biogenesis of lycopene in Mycobacterium aurum A(+), was partially digested by restriction endonucleases and generated fragments were cloned.After transformation of Escherichia coli (colorless bacteria) with the plasmids so constructed, seven orange clones were detected and found to carry the same recombinant plasmid (pC51).E. coli cells containing this plasmid synthesize neurosporene and lycopene, and were more resistant to ultraviolet irradiation than non pigmented strain.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
The plasmid pC5, which encodes biogenesis of lycopene in Mycobacterium aurum A(+), was partially digested by restriction endonucleases and generated fragments were cloned. After transformation of Escherichia coli (colorless bacteria) with the plasmids so constructed, seven orange clones were detected and found to carry the same recombinant plasmid (pC51). E. coli cells containing this plasmid synthesize neurosporene and lycopene, and were more resistant to ultraviolet irradiation than non pigmented strain.

No MeSH data available.


Related in: MedlinePlus