Science Links Japan | Cloning and Expression of Paratropomyosin Binding Connectin/Titin Domain Gene as Concerned with Meat Tenderization

Cloning and Expression of Paratropomyosin Binding Connectin/Titin Domain Gene as Concerned with Meat Tenderization

Accession number;05A1044289

Title;Cloning and Expression of Paratropomyosin Binding Connectin/Titin Domain Gene as Concerned with Meat Tenderization

Author; YAMANOUE MINORU (Kobe Univ., Fac. Agriculture, JPN) MATSUDA SUNAO (Kobe Univ., Graduate School of Sci. and Technol., JPN)

Journal Title;Shokuniku ni kansuru Josei Kenkyu Chosa Seika Hokokusho

Journal Code:X0296A

ISSN:

VOL.23;NO.;PAGE.199-204(2005)

Figure&Table&Reference;FIG.4, TBL.1, REF.9

Pub. Country;Japan

Language;Japanese

Abstract;The 43-kDa fragment digested from .BETA.-connectin/titin2 by V8 protease has been identified as the site for paratropomyosin (PTM) to bind at the A-I junction region of chicken sarcomers. Present research aimed at cloning the DNA fragment encoding connectin/titin 43-kDa fragment and its expression in E. coli DE 3 strain. The DNA fragment was amplified by nested PCR using a chicken breast muscle cDNA library and a first PCR product as first and second PCR templates respectively, and then sequenced. As the result, amino acid sequence of the 43-kDa fragment deduced from the determined cDNA showed 87.6% similarity with the sequence of A-I junction region of human cardiac muscle connectin/titin. The 43-kDa fragment was composed of four fibronectin type 3 domains and one immunoglobulin domain, and took the same domain structure as the corresponding region of human cardiac connectin/titin. For expression, DNA fragments encoding each domain of the 43-kDa fragment were amplified by PCR and cloned into pET-22b expression vectors, transfected to the E. coli. After production of recombinant domains were induced by the addition of IPTG. bands of recombinant proteins were observed on the SDS-PAGE gels in analyzing the sonicated cell lysates. (author abst.)

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