Molecular cloning of <I>Litopenaeus vannamei</I> TCP-1-eta gene and analysis on its relationship with cold tolerance

Abstract

To study the molecular mechanism of cold tolerance, we cloned TCP-1-eta homolog gene from Litopenaeus vannamei and studied its relationship with cold tolerance. Based on the sequence from electronic cloning, a pair of primers were designed, and a 1 705 bp cDNA was obtained by RT-PCR. The cDNA contains 1 629 bp ORF, which encodes a peptide of 542 aa. Tissue expression pattern by real-time PCR analysis revealed that TCP-1-eta mRNA mainly existed in muscle tissue. Cold induction by different temperatures revealed that TCP-1-eta mRNA expression started to increase after treatment at 15°C for 36 h, and the increase was remarkable after treatment at 13°C. As for treatment at 13°C for different times, the expression pattern was almost not changed during the first 36 h, but significantly increased after 36 h of treatment. A SNP (single nucleotide polymorphism) site was found in 731 bp of ORF, and the genotypes of 216 Litopenaeus vannamei were determined by PCR-RFLP. Variance analysis indicated that the SNP genotype was significantly related with cold tolerance parameter Cooling-Degree Hours (CDH), and CDH for CC type of Litopenaeus vannamei was longer than TT type.

Single nucleotide polymorphism markers with applications in aquaculture and assessment of its impact on natural populations

“…To study the molecular mechanism of cold tolerance, TCP-1-eta homolog gene cDNAwas obtained from Whiteleg shrimp Litopenaeus vannamei and was sequenced (Yin et al, 2011). TCP-1-eta gene expression depended on temperature.…”

A new ALF from Litopenaeus vannamei and its SNPs related to WSSV resistance

“…In L . vannamei , SNP 892C>T of Hsp70 was associated with resistance to Taura syndrome virus, and the level of cold tolerance was found to have an association with different genotypes of locus 731 in the ORF of TCP -1 -eta homolog gene (Zeng et al, 2008;Yin et al, 2011). Changes of the agglutinative activity against seven pathogenic bacteria were linked to mutations of three non-synonymous SNPs in the C-terminus of hemocyanin (Zhao et al, 2012).…”

Identification of cold responsive genes in Pacific white shrimp (Litopenaeus vannamei) by suppression subtractive hybridization

“…Furthermore, HSP upregulation had been associated with cold-tolerance in overwintering insects, antarctic notothenioid fish and Saccharomyces cerevisiae Parsell et al, 2013;Feder and Hofmann, 1999;Haslbeck and Vierling). While previous our lab showed that both HSP60 and four sHSPs were involved in cold tolerance in L. vannamei, the present study has implicated yet two more HSP proteins as participating in cold tolerance (Yin et al, 2011;Peng et al, 2013).…”