The Influence of Different Culture Media on the Growth and  Recombinant Protein Production of Iranian Lizard Leishmania Promastigote

Ghavidel, Afshin Abdi; Aghamiri, Seyed Mahmoud Reza; Jajarmi, Vahid; Bandehpour, Mojgan; Kazemi, Bahram

doi:10.18502/ijpa.v17i4.11282

Cited by 5 publications

(4 citation statements)

References 20 publications

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“…Multisubunit proteins have the unique three-dimensional structures of eukaryotic cells and are similar to mammalian proteins in terms of post-translational modifications ( 37 ). The production of multi-subunit proteins in Leishmania is done at a cheaper price and does not require extensive biological safety facilities, compared to mammalian cells, it has a high growth rate, its culture medium is cheap, and it grows at a temperature of 26 degrees Celsius, and does not require Incubator does not have CO2 ( 37 – 38 ).…”

Section: Discussionmentioning

confidence: 99%

Preliminary Information of Iranian Lizard Leishmania Pro-mastigote Transcriptome Sequencing by Next -Generation Sequencing (NGS) Method

Ehya,

Kalantari,

Bandehpour

et al. 2023

IJPA

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Background: A lizard Leishmania has been isolated from a lizard (Agama agilis) in Iran. Its genome sequence has not been determined, so far. Methods: The study was done at Shahid Beheshti University of Medical Sciences, Tehran, Iran in 2017-2023. Leishmania promastigotes were cultured in RPMI1640 culture medium and collected at logarithmic phase by centrigugation. Parasite RNA was extracted by the Qiagene standard kit and its quantity and quality was determined and sequenced by NGS method with Illumina PE machine at BGI Company (China). Results: The number of 8316 mRNA, 83 tRNA, 63 rRNA, 83 ncRNA, 5 snRNA, 1039 snoRNA, 36 region, and 3 repeat regions, 8343 CDS, 9597 Exon and 9292 Genes were identified in promastigote of Iranian lizard Leishmania. Conclusion: Genomic elements of Iranian lizards Leishmania (with unique characteristics) were determined and identified by NGS system.

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Section: Discussionmentioning

confidence: 99%

Preliminary Information of Iranian Lizard Leishmania Pro-mastigote Transcriptome Sequencing by Next -Generation Sequencing (NGS) Method

Ehya,

Kalantari,

Bandehpour

et al. 2023

IJPA

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“…Kazemi et al ( 6 ) isolated a strain of Leishmania , is known as Iranian Lizard Leishmania (I.L.L). This strain is non-pathogenic to mammals and offers a cost-effective option for laboratory cultivation ( 7 ). The transcriptome in the promastigote of I.L.L was identified by the NGS system ( 8 ).…”

Section: Introductionmentioning

confidence: 99%

Comparing the Yield of Recombinant Human Factor VII Protein Expressed by the rDNA-Promoter with the CMV-Promoter in Iranian Lizard Leishmania

Abdi Ghavidel,

Bandehpour,

Noori

et al. 2024

IJPA

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Background: Iranian Lizard Leishmania (I.L.L) is a nonpathogenic Leishmania strain. Due to its advantages, several recombinant proteins have been produced in this host. However, I.L.L shows a lower yield of recombinant protein expression compared to other commercial hosts. Considering the role of protease enzymes in protein digestion, we selected cysteine protease B (CPB) to investigate its impact on recombinant protein yield in I.L.L. Methods: we generated gene knockouts by utilizing homologous recombination (HR) and CRISPR methods. To assess the efficacy of the designed construct, we compared the yield of recombinant human factor VII (rhFVII) production between cells transfected with the pLEXSY-hyg2-FVII vector and the CMV-promoter-based construct (pF7cmvneo). Results: The knockout of a single CPB gene allele through the HR method or the complete knockout of all alleles through the CRISPR method led to cell death. This outcome suggests that even the deletion of a single CPB gene allele diminishes the protein to a level insufficient for the survival of I.L.L, indicating a critical dependency on the presence of this protein for the organism's viability. rhFVII exhibited a greater expression yield with the pLEXSY construct compared to the pF7cmvneo construct in I.L.L. The lower expression rate of pF7cmvneo may be influenced by epigenetic factors related to the CPC gene or the RNA polymerase used for the expression of that promoter. Conclusion: Therefore, considering alternative integration targets for CMV-promoter-based constructs and incorporating UTR sequences of I.L.L high-expression proteins in the vector may enhance recombinant protein expression rates.

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“…Furthermore, the effects of many bacterial phenotypes can be examined using LB broth medium. A previous study has shown that the growth rate of Iranian Lizard Leishmania in RPMI1640 medium and LB broth was similar, and the protein expression in LB medium was higher than that in the other three medium [ 3 ]. This indicated that LB broth medium could be a suitable medium for Iranian Lizard Leishmania culture and recombinant protein production compared with the RPMI1640 medium, brain heart infusion medium and M199 medium [ 3 ].…”

Section: Introductionmentioning

confidence: 99%

“…A previous study has shown that the growth rate of Iranian Lizard Leishmania in RPMI1640 medium and LB broth was similar, and the protein expression in LB medium was higher than that in the other three medium [ 3 ]. This indicated that LB broth medium could be a suitable medium for Iranian Lizard Leishmania culture and recombinant protein production compared with the RPMI1640 medium, brain heart infusion medium and M199 medium [ 3 ]. Another study found that the good growth without senescence of Podospora anserina was observed only on LB medium, which prolonged the health of the mycelium, while liquid cultures without senescence were observed only in LB broth medium [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

The Metabolomics Changes in Luria–Bertani Broth Medium under Different Sterilization Methods and Their Effects on Bacillus Growth

Wang,

Guo,

Chen

et al. 2023

Metabolites

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Luria–Bertani broth (LB) culture medium is a commonly used bacterial culture medium in the laboratory. The nutrient composition, concentration, and culture conditions of LB medium can influence the growth of microbial strains. The purpose of this article is to demonstrate the impact of LB liquid culture medium on microbial growth under different sterilization conditions. In this study, LB medium with four different treatments was used, as follows: A, LB medium without treatments; B, LB medium with filtration; C, LB medium with autoclaving; and D, LB medium with autoclaving and cultured for 12 h. Subsequently, the protein levels and antioxidant capacity of the medium with different treatments were measured, and the effects of the different LB medium treatments on the growth of microorganisms and metabolites were determined via 16s rRNA gene sequencing and metabolomics analysis, respectively. Firmicutes and Lactobacillus were the dominant microorganisms, which were enriched in fermentation and chemoheterotrophy. The protein levels and antioxidant capacity of the LB medium with different treatments were different, and with the increasing concentration of medium, the protein levels were gradually increased, while the antioxidant capacity was decreased firstly and then increased. The growth trend of Bacillus subtilis, Bacillus paralicheniformis, Micrococcus luteus, and Alternaria alternata in the medium with different treatments was similar. Additionally, 220 and 114 differential metabolites were found between B and C medium, and between C and D medium, which were significantly enriched in the “Hedgehog signaling pathway”, “biosynthesis of plant secondary metabolites”, “ABC transporters”, “arginine and proline metabolism”, and “linoleic acid metabolism”. LB medium may be a good energy source for Lactobacillus growth with unsterilized medium, and LB medium filtered with a 0.22 μm filter membrane may be used for bacterial culture better than culture medium after high-pressure sterilization. LB medium still has the ability for antioxidation and to keep bacteria growth whether or not autoclaved, indicating that there are some substances that can resist a high temperature and pressure and still maintain their functions.

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The Influence of Different Culture Media on the Growth and Recombinant Protein Production of Iranian Lizard Leishmania Promastigote

Cited by 5 publications

References 20 publications

Preliminary Information of Iranian Lizard Leishmania Pro-mastigote Transcriptome Sequencing by Next -Generation Sequencing (NGS) Method

Preliminary Information of Iranian Lizard Leishmania Pro-mastigote Transcriptome Sequencing by Next -Generation Sequencing (NGS) Method

Comparing the Yield of Recombinant Human Factor VII Protein Expressed by the rDNA-Promoter with the CMV-Promoter in Iranian Lizard Leishmania

The Metabolomics Changes in Luria–Bertani Broth Medium under Different Sterilization Methods and Their Effects on Bacillus Growth

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