Presentation Title

Transposon mutagenesis to identify genes involved in manganese oxidation

Faculty Mentor

Dr. Hope Johnson

Start Date

23-11-2019 8:45 AM

End Date

23-11-2019 9:30 AM

Location

72

Session

poster 2

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Manganese is an abundant transition metal involved in many species’ metabolisms, and is extensively biochemically cycled. Erythrobacter sp. SD21 is a relatively under studied marine bacteria which oxidizes manganese (Mn), and in this strain, as in others bacterial Mn oxidizing strains, the physiological function of Mn oxidation is unclear. Mn oxidation in Erythrobacter sp. SD21 is catalyzed by MopA, a heme enzyme that has been identified through protein sequencing, but other genes and proteins involved in Mn oxidation in this strain are unknown. Identifying genes involved in Mn oxidation could provide information on physiological function and on proteins that may interact with MopA. Transposon mutagenesis using pUT-miniTn5 Kmr was used to produce a library of approximately 7,000 mutants in Erythrobacter sp. SD21. Kanamycin resistance was used to select for transformants, which were then screened for changes in Mn oxidation. Sixteen slow Mn oxidizing mutants, 1 fast Mn oxidizing mutant, and 1 non Mn oxidizing mutant of Erythrobacter sp. SD21 were isolated. Growth curves of potential Mn oxidizing mutants were used to screen for mutants with general growth defects, where mutations may be in genes not directly involved in Mn oxidation. We plan to sequence the genomes of our mutants to identify the site of transposon insertion. This may lead to the identification of new genes involved in manganese oxidation in Erythrobacter sp. SD21 and a better understanding of how Mn oxidation is integrated into cellular physiology.

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Nov 23rd, 8:45 AM Nov 23rd, 9:30 AM

Transposon mutagenesis to identify genes involved in manganese oxidation

72

Manganese is an abundant transition metal involved in many species’ metabolisms, and is extensively biochemically cycled. Erythrobacter sp. SD21 is a relatively under studied marine bacteria which oxidizes manganese (Mn), and in this strain, as in others bacterial Mn oxidizing strains, the physiological function of Mn oxidation is unclear. Mn oxidation in Erythrobacter sp. SD21 is catalyzed by MopA, a heme enzyme that has been identified through protein sequencing, but other genes and proteins involved in Mn oxidation in this strain are unknown. Identifying genes involved in Mn oxidation could provide information on physiological function and on proteins that may interact with MopA. Transposon mutagenesis using pUT-miniTn5 Kmr was used to produce a library of approximately 7,000 mutants in Erythrobacter sp. SD21. Kanamycin resistance was used to select for transformants, which were then screened for changes in Mn oxidation. Sixteen slow Mn oxidizing mutants, 1 fast Mn oxidizing mutant, and 1 non Mn oxidizing mutant of Erythrobacter sp. SD21 were isolated. Growth curves of potential Mn oxidizing mutants were used to screen for mutants with general growth defects, where mutations may be in genes not directly involved in Mn oxidation. We plan to sequence the genomes of our mutants to identify the site of transposon insertion. This may lead to the identification of new genes involved in manganese oxidation in Erythrobacter sp. SD21 and a better understanding of how Mn oxidation is integrated into cellular physiology.