Judul: Konstruksi Vektor Biner dan Transformasi Gen Lisozim pada Rumput Laut Kappaphycus alvarezii Menggunakan Perantara Agrobacterium tumefaciens
Abstrak: Ice-ice disease is the biggest problem in the cultivation of seaweed Kappaphycus alvarezii. The disease is caused by bacterial infection and induced by drastic changes of water quality. Lysozyme has the ability to break down bacterial cell wall. Lysozymes are well-characterized hydrolases, which cleave beta-1,4 linkages of N-acetylglucosamine (GlcNAc) homopolymers and beta-1,4 linkages of the bacterial cell wall component GlcNAc-N-acetylmuramic acid peptidoglycan. Chicken lysozyme had lytic activities against Micrococcus lysodeikticus, Flavobacterium columnare, Aeromonas hydrophilla and Vibrio anguillarum. The purpose of this research was to construct of a binary vector pMSH1-Lis carrying chicken lysozyme (Lis) gene and introduce pMSH1-Lis on K. alvarezii. Lysozyme gene was amplified from pJfKer-Lis using PCR with specific primers for chicken lysozyme gene. Plasmid pMSH-1 and PCR product of lysozyme gene was cut with enzymes NotI and SpeI. Lysozyme gene was ligated into the expression vector pMSH1 to design a binary expression vector of pMSH1-Lis. The pMSH1-Lis was transformed to Escherichia coli DH5α by heat shock, cultured and then the plasmid was isolated. The binary vector expression was transformed into Agrobacterium tumefaciens LBA4404 by tri-parental mating. Thallus was inoculated with A. tumefaciens carrying pMSH1-Lis and then the transformed thallus was selected by adding 20 mg/L hygromycin to the culture medium.PCR analysis showed that the construction of the binary plasmid pMSH1- Lis was established. Percentage of transformation of pMSH1-Lis on K. alvarezii was 23.56%, while the efficiency of putative bud was 11.32%. PCR analysis showed that three of the regenerated thallus contained lysozyme gene. Thus, transgenic K. alvarezii was produced successfully and this can be useful for studying the mechanisms of seaweed defense against bacterial infection. Based on the percentage of the transformation, the transformation protocols need to be established for increasing transformants and regenaration thallus. Test bacterial resistance and environmental stress is necessary to examine the transformants tolerance.
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Judul: Transformasi Gen Kappa (K)-Carrageenase pada Rumput Laut Kappaphycus alvarezii
Abstrak: Foreign gene transformation technique have been applied to improve the valuable traits of farmed organism. Carrageenan content determines quality of seaweed Kappaphycus alvarezii. As the first step towards increasing of kappa(κ)-carrageenan content in K. alvarezii, in this study kappa(κ)-Carrageenase gene (κ-Car) from Pseudoalteromonas was transformed into K. alvarezii mediated by Agrobacterium tumefaciens method. The κ-Car gene involves in κ-carrageenan biosynthesis. This research consisted of three steps. The first steps of research purpose was to produce a cloning vector and an expression vector for transformation process. The κ-Car gene sequence was artificially synthesized, an then inserted in pMSH plasmid to generate a binary pMSH/κ-Car plasmid. The pMSH/κ-Car was controlled by 35S CaMV promoter (35S) and Nos terminator (tNos). Transformation of pMSH/κ-Car to Escherichia coli as cloning vector was performed using a heat-shock method, while into Agrobacterium tumefaciens was using triparental mating method. The result showed that the transformed E. coli and A. tumefaciens colonies could grow on the selective media. PCR analysis with DNA template from bacterial colonies using 35S-Forward primer (35S-F) and tNos-Reverse (tNos-R) produced DNA fragment with size of 2,000 bp, the same size with the total length of 35S promoter, κ-Car gene, and tNos sequences. Thus pMSH/κ-Car construct and A. tumefaciens transformant carrying pMSH/κ-Car were successed to be produced. The second steps of research purpose was to produce a transgenic K. alvarezii carrying κ-Car gene. Transfomation of κ-Car gene to K. alvarezii was conducted using A. tumefaciens mediation method. Analysis of K. alvarezii transgenic carrying κ-Car was performed by PCR method with two set primers. The result showed that survival percentage posttransformation of K. alvarezii explants was 36% and sprout was 88.9% while the percentage of transgenic was 100% out of the sprout explants samples. The size of DNA band of PCR product from K. alvarezii transformants using 35S-F and tNos-R set primers was 2,000 bp, the same size as PCR product of the positive control of pMSH/κ-Car plasmid as template, while no amplification product in non-transformated as negative control was found. Furthermore, PCR analysis using 35S-F and 35S-R set primers showed amplification product of 300 bp, the same size as the length of 35S promoter sequence. Thus, K. alvarezii transgenic carrying κ-Car gene had been generated..dst
Keyword: Kappaphycus alvarezii, Kappa(κ)-Carrageenase gene, transgenesis

Judul: Warna bulu kucing ras Persia : variasi pola warna dan pewarisannya
Abstrak: Kucing Persia merupakan kucing ras yang paling umum dijumpai di Indonesia. Kucing Persia memiliki bulu yang panjang dengan variasi warna bulu yang sangat beragam. Perkawinan yang lebih terarah sangat perlu dilakukan untuk meningkatkan kualitas genetik warna dan pola warna bulu kucing. Penelitian ini bertujuan untuk mempelajari variasi warna dan pola warna bulu kucing Persia yang terdapat di Indonesia, yang dilakukan di kantor pusat Cat Fancy Indonesia mulai bulan Oktober 2000 sampai Januari 2001. Data yang digunakan merupakan data pedigree kucing Persia yang tercatat di Cat Fancy Indonesia dari tahun 1997 sampai Oktober 2001 yang terdiri dari 187 jantan dan 258 betina, selain itu juga dilakukan pengamatan langsung kepada tiga penyayang dan breeder kucing yang ada di Bandung. Secara umum tercatat 104 penyayang dan breeder kucing ras di Indonesia, 74,04% terdapat di Jawa Barat dan sisanya (25%) terdapat di daerah Jakarta. Jumlah kucing Persia yang lahir di Indonesia adalah 387 kucing (87,16%), yang terdiri dari 163 (42,12%) jantan dan 224 (57,88%) betina. Jumlah kucing Persia yang diimpor ke Indonesia adalah sebanyak 57 (12,845%) yang terdiri dari 23 (40,35%) jantan dan 34 (59,65%) betina. Jumlah kucing Persia yang terdapat di Jawa Barat (84,14%) lebih banyak dibanding jumlah kucing Persia yang ada di Jakarta (14,86%). Secara umum warna bulu cream (9,01%) dan pola warna solid (34,01%) memiliki persentase tertinggi. Warna cream merupakan warna dilusi dari warna merah (red) yang dapat dihasilkan dari berbagai macam kombinasi warna bulu yang dimiliki tetua sebelumnya, sebagai contoh dari perkawinan tetua yang memiliki warna bulu cream dengan red smoke. Perkawinan antara tetua yang memiliki warna dan pola warna bulu yang tidak murni dapat menghasilkan turunan yang memiliki warna dan pola warna bulu yang bervariasi.
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