Studying and application of biotechnology in marine aquacultural species
Date Issued
2006
Date
2006
Author(s)
Chen, Hsin-Liang
DOI
en-US
Abstract
Gene delivery has become a well used technique both for economic and research purpose and been applied in aquactic organisms, such as fish, shellfish, microalgae, and macroalgae.
We microinjected the linearized plasmid pOBA-YPGHc (10 μg/μl), containing the yellowfin porgy growth hormone cDNA driven by the medaka β-actin promoter, into the testis of small abalone (Haliotis diversicolor supertexta) at a total volume of 10 μl. After polymerase chain reaction (PCR) -Southern blot detection, the gene-transfer efficiencies in larvae, juveniles, and 1-year-old adults of F1 were 90%, 92.5%, and 60%, respectively. Around 20% of F1 mature abalone contained the transgene in their gonads. Genomic Southern analysis of abalone that were PCR-Southern-positive revealed that the transgene was integrated into the genome. The average shell lengths and body weights of abalone in the gene-transfer group were significantly greater than those of the control group. We conclude that direct testis injection is a simple, high-throughput, minimally invasive, non-viral based, and efficient approach for producing transgenic line of abalone. This technique has great potential for application on many commercially important shellfish species.
Plasmid phr-YPGHc, containing the fish growth hormone (GH) cDNA driven by a heat shock protein 70A promoter and a ribulose bisphospate carboxylase small subunit 2 promoter, was transferred into the protoplast of marine microalga, Nannochloropsis oculata, by electroporation. Four transgenic clones, which the transferred phr-YPGHc was integrated into genome and stably existed at least until the 50th generation, were obtained. When we treated these transgenic microalgae by heat shock, the heterologous fish GH was produced in a amount of 0.42 to 0.27 μg /ml from the 50 ml of medium. We incubated artemia with the wild-type and transgenic N. oculata for 6 hr and then fed these microalgae-treated artemia to red-tilapia larvae. After 4-week feeding, the growth of larvae that were fed artemia incubated with transgenic microalgae was greater than that of larvae that were fed artemia incubated with non-transgenic microalgae: 239% vs 104% in weight gain, and 217% vs 91 % in body length increase, respectively. Therefore, the N. oculata enables to produce functional GH and we propose it might be an excellent bioreactor material.
Subjects
生物技術
精子
九孔
小球藻
電破
生長激素
水產養殖
sperm
abalone
Nannochlorpsis oculata
gene transfer
growth hormone
SDGs
Type
thesis
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