PERAKITAN IKAN LELE (Clarias sp.) TRANSGENIK DENGAN TEKNIK ELEKTROPORASI SPERMA

Abstract

Aplikasi teknologi transfer gen melalui perantaraan sperma dimungkinkan untuk memperbaiki kualitas genetik ikan. Keberhasilan transfer gen hormon pertumbuhan (Growth Hormone =GH) ikan budidaya melalui elektroporasi sperma dapat menghasilkan ikan transgenik. Pada penelitian ini dilakukan transfer konstruksi vektor ekspresi gen GH lele dumbo (pTarget-CMV-CgGH) ke sperma lele dumbo (strain mutiara) menggunakan kejut listrik 125 Vcm-1 dengan jumlah pulsa 3 dan 5 serta 50 Vcm-1 dengan jumlah pulsa 3 dan 5. Konsentrasi vektor yang digunakan 60 ng/?L, lama kejutan 30 milidetik dan interval kejut 0,1 detik. Dari hasil elektroporasi sperma lele mutiara sebesar 125 Vcm-1 (jumlah pulsa 3 dan 5) serta 50 Vcm-1 (jumlah pulsa 3 dan 5) ini, terdeteksi fragmen berukuran 600 bp (gen GH lele dumbo) pada sperma dan fragmen 1000 bp (96 % identik dengan GH Clarias gariepinus, no. aksesi AF416488.1) pada ikan umur 4 bulan menunjukkan keberhasilan transfer gen GH lele dumbo (GH eksogen) ke sperma lele mutiara. Rata-rata kelipatan tumbuh ikan lele mutiara transgenik umur 4 bulan (125 Vcm-1 dan 50 Vcm-1 dengan jumlah pulsa 3 dan 5) berturut- turut 3,53 kali; 3,05 kali; 2,75 kali dan 2,76 kali dari ukuran ikan non transgenik (over-ekspresi GH eksogen). GH eksogen terdeteksi pada ikan lele mutiara transgenik dan tidak terdeteksi pada ikan lele mutiara non transgenik (non elektroporasi). Ikan lele mutiara transgenik berhasildirakit dengan menyisipkan gen GH lele dumbo menggunakan teknik elektroporasi sperma.

The Quality of Ornamental Japanese Carp (Cyprinus carpio var. Koi) After Electroporation as a Gene Material Transfer Method

“…Recently, transgenesis development for aquatic animal is better to be transferred using electroporation method, because electroporation considered more suitable for the aquaculture and does not require special skills [8]. That is also seen in the efficacy of succesful gene transfer using sperm elctroporation method that has been reported to have higher efficiency compared with microinjection method, as shown in ayu fish (Plecogosus altivelis) the successful of gene transfer is 55%, in zebrafish and moad loach are 80%, in indian gold fish 25%, and 85,75 % in stripped catfish [10,11,12].…”

Genetic Relationship of the Transgenic and Non-Transgenic Mutiara Catfish with their Hybrids Sangkuriang Catfish

“…RAPD as dominant trait produced homozygotes as much as heterozygotes allele while the heterozygotes generated the polymorphic alleles (Liu, 2007). Buwono et al (2016) reported transgenic Mutiara catfish has success-Jurnal Biodjati 4(2):204-213, November 2019 http://journal.uinsgd.ac.id/index.php/biodjati fully inserted by CgGH that constructed from two GH gene derived from the fish own body (endogenous) and GH from outside the body (exogenous) thus effects of transgenic catfish revealed polymorphisms.…”

“…While on transgenesis fish, the stability of inheritance from excellent character (growth) is displayed on the F1 and F2 offspring, showing growth gene construct integrated into the fish's genome (Alimuddin et al, 2003;Rasal et al, 2016). Transgenesis application in Mutiara catfish using CgGH (Clarias gariepinus Growth Hormone), gene transfer (Buwono et al, 2016), and PhGH (Pangasius hypophthalmus Growth Hormone) gene transfer in African catfish (Dewi et al, 2013) were able to increase the catfish size two times larger than normal catfish.…”

Influence of Different Pulse Length towards Motility and Viability of Ornamental Japanese Carp (Cyprinus carpio Var. Koi) Sperm through Electroporation Method

“…Next, it was pulsed with 30 volt electric level [8], pulse length was according to treatment and the pulse number was 4 times. Post electroporation, 275 physiological NaCL (total of 300 μL solution) was added into the sperm [9]. The following step was to observe the motility and viability of the post electroporation sperm.…”