Reference Assembly of the Indian Cobra Naja Naja Provides a Comprehensive View of Its Venom Gland Genes

Reference assembly of the Indian Cobra Naja naja provides a comprehensive view of its venom gland genes Somasekar Seshagiri Diretor Associado e Cientista da equipe da Genentech, Califórnia, EUA. In India snakebites involving four venomous snake species dubbed the “big four” are responsible for over 50,000 deaths annually. Naja naja, the India cobra, is one of the “big four” venomous snakes found through out India. The genome of the Indian cobra has not been sequenced. Obtaining a well assembled genome and venom gland transcriptome will allow the identification of the complex venom components. Current anti-venoms are developed in horses and are often not effective and/or cause sever side effects. A well annotated genome will provide the template for the development of effective synthetic anti-venom antibodies with minimal side effects. Using a recently developed technique in our lab, we sequenced single chromosomes (SChrom-seq) obtained by laser capture microdissection (LCM). Using genomic DNA we generated ~60 Gb of Oxford Nanopore and ~40 Gb of PacBio single molecule long read data, and ~120 Gb of short read Illumina data. Also, we generated ~300 Gb of BioNano optical mapping data. We also generated Hi-C data. We combined these data to produce a ~1.7 Gbp assembly that contained near chromosomal assemblies. The assembled genome has an N50 of 223Mbp. In this assembly ~96% of the genome is represented by 20 scaffolds that are >1Mbp.. In addition to the genome, we sequenced RNA from nine different tissues. In particular, we sequenced RNA from venom gland using PacBio technology and combined it with short read RNA-seq to obtain full-length transcripts. We used the predicted genes from the genome and used it to assess for differentially expressed across tissues. We have identified ~150 venom gland specific genes that includes, three finger toxins, phospholipase 2, venom metallo proteases, cysteine rich small proteins, natriuretic peptides, cobra venom factor, venom kunitz type proteins, venom factor lectins, and NGF-beta..

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(Equatorial Spitting Cobra) Venom a P
The Journal of Venomous Animals and Toxins including Tropical Diseases ISSN 1678-9199 | 2011 | volume 17 | issue 4 | pages 451-459 Biochemical and toxinological characterization of Naja sumatrana ER P (Equatorial spitting cobra) venom A P Yap MKK (1), Tan NH (1), Fung SY (1) RIGINAL O (1) Department of Molecular Medicine, Center for Natural Products and Drug Research (CENAR), Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia. Abstract: The lethal and enzymatic activities of venom from Naja sumatrana (Equatorial spitting cobra) were determined and compared to venoms from three other Southeast Asian cobras (Naja sputatrix, Naja siamensis and Naja kaouthia). All four venoms exhibited the common characteristic enzymatic activities of Asiatic cobra venoms: low protease, phosphodiesterase, alkaline phosphomonoesterase and L-amino acid oxidase activities, moderately high acetylcholinesterase and hyaluronidase activities and high phospholipase A2. Fractionation of N. sumatrana venom by Resource® S cation exchange chromatography (GE Healthcare, USA) yielded nine major protein peaks, with all except the acidic protein peak being lethal to mice. Most of the protein peaks exhibit enzymatic activities, and L-amino acid oxidase, alkaline phosphomonoesterase, acetylcholinesterase, 5’-nucleotidase and hyaluronidase exist in multiple forms. Comparison of the Resource® S chromatograms of the four cobra venoms clearly indicates that the protein composition of N. sumatrana venom is distinct from venoms of the other two spitting cobras, N. sputatrix (Javan spitting cobra) and N. siamensis (Indochinese spitting cobra). The results support the revised systematics of the Asiatic cobra based on multivariate analysis of morphological characters. The three spitting cobra venoms exhibit two common features: the presence of basic, potentially pharmacologically active phospholipases A2 and a high content of polypeptide cardiotoxin, suggesting that the pathophysiological actions of the three spitting cobra venoms may be similar.
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