Mudskipper's multi-type vitellogenin Vtg gene full-length sequence and cloning method thereof

A mudskipper and protogene technology, applied in the field of molecular biology, can solve problems such as the isolation of polytype Vtg, the accuracy of detection results needs to be improved, and the mixing of polytype Vtg

Active Publication Date: 2017-06-13
SHANGHAI OCEAN UNIV +3
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  • Abstract
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  • Claims
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Problems solved by technology

However, the traditional detection method for a single type of Vtg does not isolate the corresponding protein and gene for multiple types of Vtg, so the specificity is often poor, and there are even cases whe...

Method used

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  • Mudskipper's multi-type vitellogenin Vtg gene full-length sequence and cloning method thereof
  • Mudskipper's multi-type vitellogenin Vtg gene full-length sequence and cloning method thereof
  • Mudskipper's multi-type vitellogenin Vtg gene full-length sequence and cloning method thereof

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Embodiment

[0036] This embodiment relates to a mudskipper polymorphic vitellogenin (vtg) gene, encoded protein and its application, and a method for cloning the full-length cDNA sequence.

[0037]VtgAa gene: the mudskipper polytype VtgAa gene of the present invention is characterized in that the full length of the VtgAa sequence is 5223bp, including the start codon ATG, the stop codon TAG and polyA. The open reading frame (ORF) of the gene is located at 3-5000bp, 4998bp in length, and encodes 1665 amino acids in total, in which the signal peptide is 15 amino acids. According to calculation, the molecular weight of its encoded protein is 185kDa. The encoded amino acid sequence has a typical complete vitellogenin structure: a vitellogenin domain (located at amino acid 24-594 of the encoded amino acid peptide chain), a VWFD domain (located at amino acid 1423-1555 of the encoded amino acid peptide chain) ), a polyserine region (located at amino acids 1064-1195 of the peptide chain encoding ...

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Abstract

The invention discloses mudskipper's multi-type vitellogenin Vtg gene full-length sequence and a cloning method thereof; amino acid sequences of mudskipper's vitellogenin are shown as in SEQ ID NO. 2, or in SEQ ID NO. 4 or in SEQ ID NO. 6; corresponding genes include VtgAa gene, VtgAb gene and VtgC gene. Type-III Vtg gene full-length CDSs (coding sequences) of mudskipper's Vtg family are acquired herein, the blank in mudskipper's Vtg gene family is filled, and the mudskipper's multi-type vitellogenin Vtg gene full-length sequence and the cloning method thereof are significant to the issues, such as more accurate indication for corresponding water area pollution conditions, and deeper research on the protein expression of aquatics in polluted water areas.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a full-length sequence of a mudskipper polymorphic vitellogenin (vtg) gene based on a new mudflat model fish and a cloning method thereof. Background technique [0002] Fish vitellogenin (vitellogenin, Vtg) is a yolk precursor protein synthesized by the liver under the action of estradiol (estradiol-17β, E2) and then secreted into the blood. Normally, Vtg is a female-specific serum protein that appears in the blood of female fish during yolk formation, but male fish and juveniles can also induce the generation of Vtg in vivo under the action of exogenous estrogen. [0003] The mudskipper Periophthalmus modestus is a typical representative species among the fish that live in tidal flats. Mudskippers belong to Perciformes, Goby suborders, Mudskipperidae, and Mudskipper genus. They inhabit brackish waters in estuaries, coastal tidal flats or low tide areas of bottom mud...

Claims

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Application Information

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IPC IPC(8): C07K14/46C12N15/12C12N15/10C12Q1/68
CPCC07K14/461C12N15/1096C12Q1/6851C12Q1/6879C12Q2600/158C12Q2531/113C12Q2563/107C12Q2545/101
Inventor 吴美琴陈渊戈葛珂珂练青平
Owner SHANGHAI OCEAN UNIV
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