Construction of Subclone Library of Litopenaeus vannamei Ryanodine Receptor and Determination of Its Partial Sequence

A fosmid clone containing the Litopenaeus vannamei ryanodine receptor(RyR) gene was cut by DNA shearing instrument,and the 3-5 kb fragments were collected and ligated with pUC118 HincⅡ/BAP to construct a subclone library suitable for shotgun sequencing.The titer of the library was 5.0×104 cfu·mL-1,and the recombination rate of it was 90%.The inserted fragments were in the range of 3-5 kb.One hundred positive subclones were isolated and double end-sequenced and assembled.Then a RyR gene fragment,2 550 base pairs in size,was obtained.Blast analysis showed that the deduced amino acid sequence of this fragment was highly similar to the amino-terminal sequences of Drosophila melanogaster ryanodine receptor.