scholarly journals Molecular characterization of the pollen-specific genomic clone NTPg303 and in situ localization of expression

1995 ◽  
Vol 8 (1) ◽  
pp. 11-17 ◽  
Author(s):  
K. Weterings ◽  
W. Reijnen ◽  
G. Wijn ◽  
K. van de Heuvel ◽  
N. Appeldoorn ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Genomic Clone

scholarly journals A framework for in situ molecular characterization of coral holobionts using nanopore sequencing

2020 ◽  
Author(s):  
Quentin Carradec ◽  
Julie Poulain ◽  
Emilie Boissin ◽  
Benjamin CC Hume ◽  
Christian R Voolstra ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Marker Gene ◽  
Sampling Strategy ◽  
Marker Genes ◽  
Coral Host ◽  
Sequencing Technologies ◽  
Coral Holobiont ◽  
Bacterial Microbiome

AbstractMolecular characterization of the coral host and the microbial assemblages associated with it (referred to as the coral holobiont) is currently undertaken via marker gene sequencing. This requires bulky instruments and controlled laboratory conditions which are impractical for environmental experiments in remote areas. Recent advances in sequencing technologies now permit rapid sequencing in the field; however, development of specific protocols and pipelines for the effective processing of complex microbial systems are currently lacking. Here, we used a combination of 3 marker genes targeting the coral animal host, its symbiotic alga, and the associated bacterial microbiome to characterize 60 coral colonies collected and processed in situ, during the Tara Pacific expedition. We used Oxford Nanopore Technologies to sequence marker gene amplicons and developed bioinformatics pipelines to analyze nanopore reads on a laptop, obtaining results in less than 24 hours. Reef scale network analysis of coral-associated bacteria reveals broadly distributed taxa, as well as host-specific associations. Protocols and tools used in this work may be applicable for rapid coral holobiont surveys, immediate adaptation of sampling strategy in the field, and to make informed and timely decisions in the context of the current challenges affecting coral reefs worldwide.

scholarly journals Characterization of two novel Microplitis demolitor polydnavirus mRNAs expressed in Pseudoplusia includens haemocytes

2000 ◽  
Vol 81 (12) ◽  
pp. 3049-3058 ◽  
Author(s):  
D. Trudeau ◽  
R. A. Witherell ◽  
M. R. Strand
Keyword(s):  
Amino Acids ◽  
Northern Blotting ◽  
Genomic Clone ◽  
Pseudoplusia Includens ◽  
Microplitis Demolitor ◽  
Multiple Copies ◽  
Epidermal Growth ◽  
Braconid Wasp

The braconid wasp Microplitis demolitor carries M. demolitor polydnavirus (MdPDV) and parasitizes the larval stage of the moth Pseudoplusia includens. M. demolitor injects MdPDV into P. includens larvae when it lays an egg and the virus infects various cells including haemocytes. Two new MdPDV transcripts expressed in host haemocytes were characterized in this study. Screening of an MdPDV-infected haemocyte cDNA library identified a 0·4 kb cDNA encoding a predicted protein of 103 amino acids which was named Egf0·4. This protein contained a cysteine-rich epidermal growth factor (EGF)-like motif at its N terminus that was similar to the EGF-like domains in the previously identified MdPDV genes egf1·5 and egf1·0. Sequencing of the genomic clone pMd-10 indicated that it contained the egf0·4 gene, which consisted of two introns and three exons. This gene was located on MdPDV segment O and appeared to exist in multiple copies. A nucleic acid and expression screen identified a 1·8 kb cDNA encoding a predicted protein of 515 amino acids designated Glc1·8. This protein consisted of a heavily glycosylated central core of six tandemly arranged repeats flanked by hydrophobic N- and C-terminal domains. Northern blotting and in situ hybridization studies indicated that both egf0·4 and glc1·8 were expressed in MdPDV-infected host haemocytes. Immunocytochemical studies also indicated that Glc1·8 localized to the cell surface.

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