scholarly journals Molecular Cloning and Sequence Analysis of the cDNAs Encoding Toxin-Like Peptides from the Venom Glands of Tarantula Grammostola rosea

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Tadashi Kimura ◽  
Seigo Ono ◽  
Tai Kubo
Keyword(s):  
Sequence Analysis ◽  
Molecular Cloning ◽  
Real Time ◽  
Cdna Library ◽  
Real Time Pcr ◽  
Bioactive Peptides ◽  
Venom Gland ◽  
Unique Peptide ◽  
Cdna Sequences ◽  
Venom Glands

Tarantula venom glands produce a large variety of bioactive peptides. Here we present the identification of venom components obtained by sequencing clones isolated from a cDNA library prepared from the venom glands of the Chilean common tarantula, Grammostola rosea. The cDNA sequences of about 1500 clones out of 4000 clones were analyzed after selection using several criteria. Forty-eight novel toxin-like peptides (GTx1 to GTx7, and GTx-TCTP and GTx-CRISP) were predicted from the nucleotide sequences. Among these peptides, twenty-four toxins are ICK motif peptides, eleven peptides are MIT1-like peptides, and seven are ESTX-like peptides. Peptides similar to JZTX-64, aptotoxin, CRISP, or TCTP are also obtained. GTx3 series possess a cysteine framework that is conserved among vertebrate MIT1, Bv8, prokineticins, and invertebrate astakines. GTx-CRISP is the first CRISP-like protein identified from the arthropod venom. Real-time PCR revealed that the transcripts for TCTP-like peptide are expressed in both the pereopodal muscle and the venom gland. Furthermore, a unique peptide GTx7-1, whose signal and prepro sequences are essentially identical to those of HaTx1, was obtained.

scholarly journals Characterization and Localization of Calb2 in Both the Testis and Ovary of the Japanese Flounder (Paralichthys olivaceus)

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1503
Author(s):  
Yuting Xiang ◽  
Yahui Wu ◽  
Haoran Zhang ◽  
Jikui Wu ◽  
Junling Zhang
Keyword(s):  
Real Time ◽  
Western Blotting ◽  
Real Time Pcr ◽  
Paralichthys Olivaceus ◽  
Neighboring Gene ◽  
Syntenic Relationship ◽  
Reading Frame ◽  
Cdna Sequences ◽  
Its Gene ◽  
And Function

Although its function in mammalian gonads has been gradually recognized, the expression and function of calretinin (CALB2)—a Ca2+-binding protein—in the testis and ovary of fish are still unclear. Here, we identified the cDNA sequences of calb2 in Paralichthys olivaceus (P. olivaceus); analyzed its gene structure and phylogenetic and syntenic relationship by bioinformatics; and investigated its tissue distribution and localization in the gonads by real-time PCR, western blotting, and immunohistochemistry. The P. olivaceuscalb2 gene has 11 exons and 10 introns, and the full-length cDNA is 1457 bp, including an open reading frame (ORF) of 816 bp encoding 271 amino acids. The CALB2 of P. olivaceus has a higher homology with Lates calcarifer (99%) compared with other species. The conserved synteny of calb2 neighboring gene loci was also detected in fish. Real-time PCR showed that the expression of calb2 mRNA is abundant not only in the brain, but also in the gonads, and exhibits a higher expression in the testis than in the ovary. Western blotting indicated that the CALB2 protein has a higher expression in the testis compared with the ovary. Immunohistochemistry demonstrated that the CALB2 protein appears in Leydig cells and the ovarian germ epithelium. These results reveal that calb2 plays an important role in the gonads of P. olivaceus.

Molecular cloning and nucleotide sequence analysis of genes from a cDNA library of the scorpion Tityus discrepans

Biochimie ◽  
2009 ◽  
Vol 91 (8) ◽  
pp. 1010-1019 ◽  
Author(s):  
Gina D'Suze ◽  
Elisabeth F. Schwartz ◽  
B.I. García-Gómez ◽  
Carlos Sevcik ◽  
Lourival D. Possani
Keyword(s):  
Sequence Analysis ◽  
Nucleotide Sequence ◽  
Molecular Cloning ◽  
Cdna Library ◽  
Nucleotide Sequence Analysis

scholarly journals OCCURRENCE OF TICK-BORNE PATHOGENS IN STRAY DOGS FROM HAVANA, CUBA

2018 ◽  
Vol 3 (4) ◽  
pp. 158-159
Author(s):  
B. G. Corona ◽  
A. A. Díaz-Sánchez ◽  
M. L. Meli ◽  
E. V. Cañizares ◽  
L. R. Arias ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
16S Rrna ◽  
Real Time ◽  
Real Time Pcr ◽  
Ixodid Tick ◽  
Ehrlichia Canis ◽  
Hepatozoon Canis ◽  
Anaplasma Platys ◽  
Rickettsia Felis ◽  
Stray Dogs

The article presents the results of examination of stray dogs from Havana, Cuba for six ixodid tick-borne diseases. Analysis was carried  out using real-time PCR. Overall 107 dogs, 95 (89.09 %) were infected.  41 dogs (38.31 %), 66 (61.68 %), 28 (26.17 %) and 40 (37.38 %)  were found to be infected with Anaplasma platys, Ehrlichia canis,  Rickettsia spp. and Hepatozoon canis, respectively. Sequence analysis of 16S rRNA and groEL genes for Rickettsia spp. revealed 99 % identity  with Rickettsia felis. There were no dogs infected with A. phagocytophilum and Borrelia spp.

Molecular Cloning of Disintegrin-like Transcript BA-5A from a Bitis arietans Venom Gland cDNA Library: A Putative Intermediate in the Evolution of the Long-Chain Disintegrin Bitistatin

2006 ◽  
Vol 63 (1) ◽  
pp. 142-152 ◽  
Author(s):  
Paula Juárez ◽  
Simon C. Wagstaff ◽  
Jenny Oliver ◽  
Libia Sanz ◽  
Robert A. Harrison ◽  
...  
Keyword(s):  
Molecular Cloning ◽  
Cdna Library ◽  
Venom Gland ◽  
Long Chain

Sequence analysis and gene amplification study of the penicillin biosynthesis gene cluster from different strains of Penicillium chrysogenum

Biologia ◽  
2010 ◽  
Vol 65 (1) ◽  
pp. 1-6 ◽  
Author(s):  
Roman Šmidák ◽  
Martina Kralovičová ◽  
Beatrica Ševčíková ◽  
Mária Jakubčová ◽  
Ján Kormanec ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Real Time ◽  
Real Time Pcr ◽  
Penicillium Chrysogenum ◽  
Gene Dosage ◽  
Single Copy ◽  
Nucleotide Sequence Analysis ◽  
Penicillin Biosynthesis ◽  
Biosynthesis Gene Cluster ◽  
Genomic Changes

AbstractIndustrial strains of Penicillium chrysogenum possess many genomic changes leading to higher levels of penicillin. In this work several production and wild-type strains of Penicillium chrysogenum were used in comparative nucleotide sequence analysis of the biosynthesis cluster. The alignments confirmed sequence conservation not only in promoter regions of the biosynthesis genes but also throughout the entire 44.7-kbp genomic fragment comprising the whole biosynthesis cluster with 15.5-kbp and 13.1-kbp flanking regions. As another titre-enhancing mechanism we subsequently examined gene dosage in two production strains introduced here, NMU2/40 and B14. Quantitative real-time PCR and Southern blot analysis showed the amplification of the biosynthesis genes in both these strains. Through the real-time PCR method the exact copy number was estimated for each of the pcbAB, pcbC and penDE genes. The equal pool of all three genes per genome was confirmed for the both production strains indicating that in these strains the entire penicillin cluster has been amplified as an intact element. Penicillium chrysogenum NMU2/40 was found to carry four copies of the cluster, while six copies were estimated for B14. This also proves the contribution of the additional titre-enhancing mechanisms in both strains, since the industrial data referred much higher production of these strains compared with the single copy reference strain NRRL 1951.

Molecular cloning of novel serine proteases and phospholipases A2 from green pit viper (Trimeresurus albolabris) venom gland cDNA library

Toxicon ◽  
2006 ◽  
Vol 47 (3) ◽  
pp. 279-287 ◽  
Author(s):  
Ponlapat Rojnuckarin ◽  
Chuanchom Muanpasitporn ◽  
Lawan Chanhome ◽  
Jaradpong Arpijuntarangkoon ◽  
Tanin Intragumtornchai
Keyword(s):  
Molecular Cloning ◽  
Cdna Library ◽  
Serine Proteases ◽  
Venom Gland ◽  
Phospholipases A2

scholarly journals Prevalence and Abundance of Latently Transcribed Varicella-Zoster Virus Genes in Human Ganglia

2006 ◽  
Vol 81 (6) ◽  
pp. 2950-2956 ◽  
Author(s):  
Randall J. Cohrs ◽  
Donald H. Gilden
Keyword(s):  
Sequence Analysis ◽  
Varicella Zoster Virus ◽  
Real Time ◽  
Real Time Pcr ◽  
Trigeminal Ganglia ◽  
Mrna Transcript ◽  
Varicella Zoster ◽  
Virus Latency ◽  
Viral Genes ◽  
Latently Infected

ABSTRACT In human ganglia latently infected with varicella-zoster virus (VZV), sequence analysis has revealed that five viral genes (VZV genes 21, 29, 62, 63, and 66) are transcribed. However, their comparative prevalence and abundance are unknown. Here, using real-time PCR, we analyzed 28 trigeminal ganglia from 14 humans for RNA corresponding to the five virus genes known to be transcribed in latently infected human ganglia. The most prevalent transcript found was VZV gene 63 (78%), followed by gene 66 (43%), gene 62 (36%), and gene 29 (21%). No gene 21 transcripts were detected in any of the 28 ganglia. VZV gene 63 RNA was also the most abundant (3,710 ± 6,895 copies per 1 μg of mRNA) transcript detected in latently infected human ganglia, followed by VZV gene 29 (491 ± 594), VZV gene 66 (117 ± 85), and VZV gene 62 (64 ± 38). Thus, the repeated detection and high abundance of VZV gene 63 transcripts in latently infected ganglia suggests that VZV gene 63 may be more important for the maintenance of virus latency than the less abundantly transcribed and randomly detected VZV genes 21, 29, 62, and 66.

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