Judul: Gene Expression Analysis of Heat Shock Protein-70 in Giant Gourami after Transportation Stress
Abstrak: Respons fisiologis pada tubuh ikan yang terpapar stresor salah satunya adalah sintesis heat shock protein-70 (HSP70). Sintesis HSP70 merupakan respons dari sel yang berfungsi menjaga kelengkapan protein sel, pemindahan protein, menjaga dari pelipatan prematur pada sel dan mencegah penumpukan protein pada sel. Penelitian ini bertujuan untuk menganalisis tingkat ekspresi gen HSP70 pada benih ikan gurami yang terpapar stres transportasi. Penelitian dilakukan dengan mentransportasikan ikan perlakuan (3 ulangan) dengan plastik kemas berukuran 40 × 60 cm2 berisi ikan dengan panjang 6,02±0,62 cm dan bobot 8,28±0,49 g dengan kepadatan 40 ikan per-plastik. Ikan kontrol (1 ulangan) dikemas dengan spesifikasi yang sama seperti ikan perlakuan dan diletakkan dalam akuarium tanpa ditransportasikan. Ikan kemudian langsung dibedah pascatransportasi dan diambil sampel hati (liver) untuk ekstraksi RNA dan kemudian disintesis menjadi cDNA. Sampel cDNA lalu diamplifikasi dengan metode PCR menggunakan primer HSP70 dan β-aktin. Tingkat ekspresi gen ditentukan menggunakan metode PCR semikuantitatif. Hasil elektroforesis menunjukkan bahwa ukuran pita pada sampel primer HSP70 sebesar 267 bp dan primer β-aktin sebesar 192 bp. Analisis ekspresi gen HSP70 pada ikan pasca paparan stres transportasi menunjukkan bahwa tidak ada perbedaan nyata antara ikan kontrol, rentan dan tahan stres setelah dilakukan uji statistik (P>0,05). Stres transportasi tidak memberikan pengaruh yang signifikan terhadap ekspresi gen HSP70 pada hati ikan gurami., One of the physiological responses of fish exposed to stress transportation is the synthesis of heat shock protein-70 (HSP70). Synthesis of HSP70 is a cell response to stressors that function to maintain the completeness of cell proteins, transfer proteins, prevent premature folding of cells, and prevent protein accumulation in cells. This research aimed to analyze the expression level of the HSP70 gene in giant gourami fry exposed to transportation stress. The research was conducted by transporting treated fish (3 replicates) with plastic containers measuring 40 × 60 cm2 containing fish with a length of 6.02±0.62 cm and a weight of 8.28±0.49 g and a density of 40 fish per plastic. Control fish (1 replicate) were packed with the same specifications as the treatment fish and placed in the aquarium without being transported. The fish were then dissected after transport and liver samples were taken for RNA extraction and then synthesized into cDNA. The cDNA samples were then amplified by the PCR method using HSP70 and β-actin primers. Gene expression level was determined by using the semi-quantitative PCR method. Electrophoresis results showed that the band size in the HSP70 primer sample was 267 bp and the β-actin primer was 192 bp. HSP70 gene expression analysis in giant gourami after being exposed to transportation stress showed that there was no significant difference between control, prone to-, and stress-resistant fish after statistical tests (P>0.05). Transport stress did not have a significant effect on the expression of the HSP70 gene in giant gourami liver.
Keyword: Giant Gourami, HSP70, Primer, Stress, Transportation