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Molecular Cloning cDNAs and Bioinformation Analysis of Follicle-stimulating Hormone (FSH)-β Subunit and Luteinizing Hormone (LH)-β Subunit of Schizothorax Prenanti

Author: CaoHongTao
Tutor: MaHengDong；ZengChangJun
School: Sichuan Agricultural University
Course: Animal Genetic Breeding and Reproduction
Keywords: To mouth together schizothoracine fish Follicle-stimulating hormone Luteinizing hormone cDNA clones Rapid Amplification of cDNA ends
CLC: S917.4
Type: Master's thesis
Year: 2010
Downloads: 119
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Abstract


Gonadotropin (GTH) is the vertebrate brain anterior pituitary cells synthesize and secrete a glycoprotein hormone, the main physiological role is to stimulate the growth of germ cell development, mature, normal ejaculation and ovulation. The the fish gonadotropin GTH I and GTH II respectively, corresponds to mammals which follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Gonadotropin is composed of two alpha and beta subunits, and is connected to the non-covalent bond to form a dimer structure, wherein the alpha subunit amino acid composition of the same, distinct p subunit amino acid composition having specific biological learn the role. FSHβ subunit and the LHβ subunit structure and function so the research will help to understand the physiological condition of the livestock and poultry, fish reproductive, and of great significance to further improve the reproductive efficiency of animals, and tap the potential of animal breeding. Sarkozy mouth Schizothorax (Schizothorax prenanti) as experimental materials used in this study, total RNA was extracted from Qi mouth schizothoracine the fish pituitary gland. Design for FSHβ subunit Qi mouth Schizothorax Hormone Beta subunit gene-specific primers, rapid amplification of cDNA ends (RACE) was cloned Qi the mouth Schizothorax the cDNA sequence, and the sequence to the correlation analysis. The results showed that: (1) Qi mouth schizothoracine fish FSHβ subunit cDNA sequence length of 624bp, and the length of the sequence of the coding region (CDS) of 393 bp encoding 130 amino acids; Hormone Beta subunit cDNA sequence length of 640bp coding region (CDS ) a sequence length of 441bp, encoding 146 amino acids. Bioinformatics analysis showed that Qi mouth Schizothorax FSHβ subunit is a negatively charged protein has a transmembrane helix and three major hydrophobic regions with molecular weight of 14.51kDa, pI of 5.28. Its amino acid sequence having 12 phosphorylation sites; LHP subunit is also a protein having 1 transmembrane helix and four major hydrophobic regions with a negative charge, and a molecular weight of 16.34kDa and pI of 4.98. Its amino acid sequence having 11 phosphorylation sites. (2) Qi mouth schizothoracine fish and other vertebrates, including mammals, birds, reptiles, amphibians, fish and other species two genes encoding amino acid homology analysis showed: FSHP subunit Cypriniformes carp, the highest homology of 96.9%, the Japanese toad amphibian lowest homology of 37.6%; LHP subunit homology with carp highest, reaching 93.1%, 43.2% and the lowest of mankind. Homology Show: Hormone Beta subunit same carp highest homology Qi mouth schizothoracine the fish FSHβ subunit carp of homologous up 96.2% the Japanese toad minimum and amphibians to 46.9%; 95.8% with birds Japanese quail minimum of 50.5%. The preliminary findings between the two sequences can be determined clone for Qi mouth schizothoracine the fish FSHβ subunit and LHβ subunit. (3) the use of MAGA4.0 NJ phylogenetic trees and MP phylogenetic tree constructed based on the amino acid sequences of 22 species of vertebrates that: Qi mouth schizothoracine the fish is a Cypriniformes, form a cluster with carp, grass carp, herring, to keep with the traditional taxonomy consistent similarities of the degree of genetic distance and systems branch Show FSHp subunit evolutionary rate relatively conservative Hormone Beta subunit in birds, reptiles, mammals evolved faster than other vertebrates. Can reveal the evolution of the two glycoprotein gene and its function appears accompanied by the evolution of species.

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CLC: > Agricultural Sciences > Aquaculture, fisheries > Aquatic basic science > Aquatic Biology > Aquatic Zoology
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