Study On The Myofibril Protein And Its Gel Properties Of Squid

Although Cyprinus carpio wuyuanensis, Gaohan Carp Cultivation and other fine varietieswith growing fast, low temprature tolerance and strong characteristic had been cultivated in thenorthern region through traditional cross-breeding and selection of breeding methods, most of thethermophilic fishes such as mud carp (Cirrhinus molitorella)、tilapia, etc. can’t rely onconventional breeding methods to obtain cold strain. This requires us to research the lowtemperature mechanism of fish in gene expression levels. This experiment is to study the mainexpression of cold-resistance gene MIPS that come from the northeast area with low temperatureresistance traits of Heilongjiang carp (Cyprinus carpio) in zebrafish and Tilapia.The results of exogenous expression vector fluorescent observation in zebrafish showed thatboth the recombinant expression vector do not affect the expression of EGFP, just expressingintensity, the EGFP gene expression of recombinant expression vector that do not insert theexogenous gene is higher than the one with insertion. The successful construction of recombinantexpression vector displays goldfish Tgf2transposon and regular expression vector can mediateexogenous gene expression in zebrafish.The PCR results of transgenic zebrafish F1, F2generation and tilapia F1generation showedthat the coding region of the target gene MIPS can completely integrate into the genome of thezebrafish and tilapia, detecting the integration efficiency of F1transgenic zebrafish that injectedpTgf2-EF1α-MCS-EGFP expression vector up to31.4%; the integration efficiency of F1transgenic zebrafish that injected common expression vector up to30.4%; the integrationefficiency of F2transgenic zebrafish that injected pTgf2-EF1α-MIPS-EGFP expression vector upto18.1%; the integration efficiency of tilapia that injected pTgf2-EF1α-MIPS-EGFP expressionvector up to6.01%, the results confirmed that exogenous gene of MIPS are express-ed intransgenic zebrafish and tilapia.The results of low temperature tolerance experiment that used the PCR test to detect thetransgenic zebrafish genes F2individual and tilapia F1individual showed that the transgenic grouphalf lethal time is higher than non transgenic control group. According to the chi square calculationformulas that compare between the two groups of half lethal time, X2=0.000is less than0.05, thedifference between the transgenic and control groups in half lethal time is not significant, thetransgenic individuals and control one do not differ significantly in the low temperature toleranceability. Zebrafish real-time PCR experiment showed that the gene MIPS in room temperaturetransgenic zebrafish expressed the highest. The expression of transgenic zebrafish that finally died in the low-temperature tolerance test is followed; The tilapia real-time PCR results showed that thehighest expression of cold MIPS gene is in the spleen, followed by gill, intestine, kidney, heart andliver.