title: Isolation of Tanichthys albonubes beta actin gene and production of transgenic Tanichthys albonubes.
abstract: A beta actin cDNA of Tanichthys albonubes was isolated through the RT-PCR and RACE approach. The cDNA was 1,787-bp in length, including a 1,128-bp CDS, a 95-bp 5'UTR and a 564-bp 3'UTR. Genomic DNA containing the transcription region and 5'-flanking region was cloned based on the beta actin cDNA by Genome walker. A 3,000-bp beta actin gene promoter was then produced by PCR according to the sequences of the 5'-flanking region and the first intron. This promoter consisted of a 1,800-bp 5'-flanking region, and a 1,200-bp 5'-UTR. 3 transcription elements, CAAT box, CArG motif and TATA box were found in the 5'-flanking region. This promoter was inserted into the vector pDsRed2-1 and microinjected into fertilized eggs of Tanichthys albonubes to prove its transcription activity. The beta actin promoter and GH CDS of Tanichthys albonubes were then fused to construct an expression vector pTLA-GH. GH-transgenic Tanichthys albonubes was obtained by microinjection of the pTLA-GH into the fertilized eggs. Fast-growth individuals were observed in the transgenic group and the body weight of the largest individual was 2.1-fold that of the maximum in its non-transgenic siblings in 100 dph. In addition, a co-injection strategy was employed with pTLA-DsRed and pTLA-GH vector and proven to enhance the efficiency of GH-transgenic fish detection.
authors.affiliation: College of Aqua-life Science and Technology, Shanghai Ocean University, Shanghai 200090, China.
authors.forename: Er-meng
authors.initials: EM
authors.lastname: Yu
authors.forename: Xing
authors.initials: X
authors.lastname: Ye
authors.forename: Hai-ying
authors.initials: HY
authors.lastname: Wang
authors.forename: Jun-jie
authors.initials: JJ
authors.lastname: Bai
authors.forename: Shi-ling
authors.initials: SL
authors.lastname: Xia
authors.forename: Hai-hua
authors.initials: HH
authors.lastname: Lao
authors.forename: Qing
authors.initials: Q
authors.lastname: Jian
mesh_terms: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D000199:Actins / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D003001:Cloning, Molecular; D022801:Complementarity Determining Regions; D003530:Cyprinidae / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D029941:Fish Proteins / Q000235:genetics*; D005822:Genetic Vectors; D013006:Growth Hormone / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D000199:Actins; D022801:Complementarity Determining Regions; D017931:DNA Primers; D018076:DNA, Complementary; D029941:Fish Proteins; D008164:Luminescent Proteins; D011994:Recombinant Proteins; C413663:fluorescent protein 583; D013006:Growth Hormone
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