Analysis Of A CDNA Library Constructed By Low Temperature Treated Young Litopenaeus Vannamei And Two Functional Genes

A muscle cDNA library of Litopenaeus vannamei which aged 2-3 months under Low temperature stress was constructed with the SMARTTM cDNA Library Construction Kit.The titer of optimal primary library was 1.05�106 pfu/mL.The percentages of the recombinant clones of primary and amplified libraries were over 94%.The insert sizes were longer than 400 bp with an average of 1000 bp.A positive clone containing three alternative splicing was sequenced and identified encoding Troponin I gene.This library provided a useful resource for the functional genomic research of Litopenaeus vannamei.We get the full sequence of Litopenaeus vannmei Metallothionein gene from the Subtractive of low temperature processing Litopenaeus vannmei's liver and pancreas and the cDNA library of low temperature processing young Litopenaeus vannmei.The complete gene sequence was consisting of 425 nucleotides,encoding corresponding 58 amino acids.The Open reading frame of the sequence has 177 bp,Non-coding region in the upper reaches of the firsr start codon(ATG)has 98 bp and there is 150 bp in the downstream of termination codon.The sequence of amino acids was rich in cysteine and Cys-X(1-3)-Cys structure which is Mt classic struchture.Sequence alignment analysis showed that the homology of Litopenaeus vannmei Metallothionein and Homarus americanus was up to 72.4%.The quantification result of Litopenaeus vannmei Metallothionein mRNA showed that the expression Litopenaeus vannmei Metallothionein gene gets the highest levels in the ovarian and the expression was found to be high level in the ovarian stages of development.Furthermore,the expression of the low temperature processing Litopenaeus vannmei's liver and pancreas is up trend.To conclude,the results and data above,grounded on the experiment of Litopenaeus vannmei Metallothionein gene,might offer essential information and material in development and research about ovarian function and low temperature stress.