ISOLATION AND IDENTIFICATION OF PARTIAL GROWTH HORMONE (GH) mRNA OF TIGER SHOVELNOSE CATFISH Pseudoplatystoma fasciatum (LINNAEUS, 1766)

Q4 Environmental Science Indonesian Aquaculture Journal Pub Date : 2022-12-30 DOI:10.15578/iaj.17.2.2022.107-119
E. P. Hayuningtyas, Eni Kusrini, A. Abinawanto
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Abstract

The growth of fish is regulated by growth hormones secreted in a limited amount by its pituitary glands. Tiger shovelnose catfish has a relatively faster growth indicating that some growth hormones suspectedly play roles in the process and could be used in improving other farmed fish species. This study aimed to isolate and identify the growth hormone (GH) mRNA gene in Tiger shovelnose catfish (Pseudoplatystoma fasciatum Linnaeus, 1766). The sample was isolated from the pituitary, the fish measuring 602 g and 43 cm body length; Total RNA was extracted using the Tri Reagent Kit, followed with cDNA synthesis. The success of the extraction was confirmed by quantification and PCR using the β-actin gene as an internal control. GH mRNA gene was isolated by RT-PCR method, with degenerated primers from seven catfish species sequence data in the NCBI gene bank. The single band from GH gene was cloned and sequenced. Total RNA quantification with a concentration of 227 ng/µL and purity of 1.821. The successful isolation of mRNA from the pituitary gland was confirmed by amplifying the β-actin gene generated at 300 bp. This isolation of the GH mRNA gene had a sequence length of 234 bp. Tiger shovelnose catfish GH gene consists of 17 amino acid residues. The GH gene of Tiger shovelnose catfish was close to that of striped catfish (P. hypophthalmus) and Indian catfish (Clarias batrachus) with almost similar homology value of 90.6 %. Partial GH gene from tiger shovelnose catfish can be used a molecular marker in revealing the role of growth hormone on fish development, fish biology, growth gene expression, selective breeding and other mechanisms related to the aquaculture.
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虎舌状鲶鱼(LINNAEUS,1766)部分生长激素(GH)mRNA的分离与鉴定
鱼的生长受脑下垂体分泌的有限数量的生长激素的调节。虎铲鼻鲶鱼的生长速度相对较快,这表明一些生长激素可能在这一过程中起作用,并可用于改善其他养殖鱼类。本研究旨在分离和鉴定虎铲鼻鲶鱼(Pseudoplatystoma fasciatum Linnaeus, 1766)生长激素(GH) mRNA基因。样品从垂体中分离出来,鱼长602 g,体长43 cm;用Tri Reagent Kit提取总RNA,合成cDNA。以β-肌动蛋白基因为内参,通过定量和PCR验证提取成功。利用NCBI基因库中7种鲶鱼序列数据的退化引物,采用RT-PCR方法分离出生长激素mRNA基因。克隆了生长激素基因的单条带,并对其进行了测序。总RNA定量,浓度为227 ng/µL,纯度为1.821。通过扩增300 bp处生成的β-肌动蛋白基因,证实成功分离垂体mRNA。该基因的序列长度为234bp。虎铲鼻鲶鱼生长激素基因由17个氨基酸残基组成。虎铲鼻鲶鱼的生长激素基因与条纹鲶鱼(P. hypophthalmus)和印度鲶鱼(Clarias batrachus)的生长激素基因同源性接近,同源性为90.6%。虎铲鼻鲶鱼部分生长激素基因可作为一种分子标记,揭示生长激素在鱼类发育中的作用、鱼类生物学、生长基因表达、选择育种等与水产养殖相关的机制。
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来源期刊
Indonesian Aquaculture Journal
Indonesian Aquaculture Journal Environmental Science-Water Science and Technology
CiteScore
0.50
自引率
0.00%
发文量
9
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