The Use of Cytochrome B Gene as a Specific Marker of the Rat Meat (Rattus Norvegicus) on Meat and Meat Products

Falsification of the origin of livestock meat and its processed with rat meat is a problem that must be overcome to ensure food safety. One way that is often used to detect forgeries by using cytochrome b gene as a marker. The purpose of this study was to create a specific primer derived from cytochrome b sequences in rat (Rattus norvegicus) as the DNA marker to detect any contamination of rat meat on fresh livestock meat and its processed meat products. Meatballs were made from beef meat with the addition of rat 1%-25%, and the meatballs were obtained from traditional markets. DNA extraction was conducted from seven species (goat, chicken, cattle, sheep, pig, horse, and rat) by using phenol-chloroform. The highest success rate in detecting the presence of rat meat in a mixture of beef meatballs at concentration of 15% was 100%. The specific fragment of cytochrome b gene in R. norvegicus has no similarity with the cytochrome b gene from six other species, so it can be used as molecular markers to detect the presence of rat meat contamination in the processed of meat products. Amplified fragment length for goats, chickens, cattle, sheep, pigs, horses, and rats 157, 227, 274, 331, 398, 439 and 603 bp respectively. The amplification of cytochrome b gene in seven species of animals with different fragment length indicated the specificity of cytochrome b gene sequences among species.

ensure food safety; and (ii) traceability of individuals to their source breed or species to detect possible labelling adulteration (Dalvit et al., 2007). The process of determination of the existence of fraud or other species contamination in a product (Zhang et al., 2007;Ghovvati et al., 2009). This method uses a universal DNA marker (2004) has been reported that spe-OE' OE1 oeŽšžŽ-OEŽoe1 Porcine Repetitive Element 1 (PRE-1) can be used to detect the pigs (Sus scrofa) and their relatives. et al., 2007).
Cyt b gene is a gene that is often used to compare multiple phylogenetic species in the same genus or family, the diversity of the cyt b gene has been used , sheep (Ovis Aries), goats (Capra hircus), roe buck (Capreolus capreolus) and red deer (Cervus elaphus) (Wolf et al., 1999). Species of rat ( Š4žoe -˜›ŸŽ•'OEžoe) have long oeŽšžŽ-OEŽoe1˜•1 cyt b gene of 1143 bp (Naidu et al., 2010), while in Bos taurus 1140 bp in length (Geng and Chang, 2008). Research report on the contamination of livestock meat and processed meat products with rat meat in used as a mixture of DNA to detect the presence of rat meat in livestock raw meat and meat products. The Matsunaga et al. (1999). Forward primer used for the seven animals were the same, namely

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Blood samples taken from goat, chicken, cow, sheep, pig, horse, and rat, and meatballs from meat beef mixed with rat meat were used for DNA extraction. There were two kinds of meatballs used in this experi- beef with rat meat with a composition ratio of 1%, 5%, 10%, 15%, and 25%, and the second was purchase directly from traditional markets in Bogor. Rat used as a compound derived from the species ï -˜›ŸŽ•'OEžoe albino strain. The process of DNA isolation used phenol-chloroform method (Sambrook & Russell, 2001). Sampling from traditional market was conducted to determine measurement of DNA purity and its concentration was done by using a spectrophotometer to ensure successful
1™›˜•žOEŽ•ï1 According to Nuraini (2004), the use of spices and other ingredients in the product meatballs, such as starch and sodium tripolyphosphate, will cause the DNA extracted compound still mixed with contaminants, such as polysaccharides, oligopeptide, and other organic materials. The process of extraction methods determined origin of the material extracted. Generally, the raw tissue was easier to extract than the tissue that had been cooked or mixed with other ingredients. The DNA concentration used in the process of copying DNA through PCR was 50 ug/ml. DNA samples with concentrations higher than 50 ug/ml could be processed by adding destilated water. Matsunaga et al. (1999) for six species of livestock, pig, and horse according to Matsunaga et al. (1999) were 157, 227, 274, 331, 398 and 439 bp, respectively. In
The highest success rate in detecting the presence of rat meat in a mixture of beef meatballs at concentration ˜•1 W[-1 Šoe1 WVV-ï1 'Ž1 oe™ŽOE' OE1 •›Š•-Ž-•1˜•1 cytochrome b gene in ï -˜›ŸŽ•'OEžoe has no similarity with the cytochrome b gene from six other species, so it can be used as molecular markers to detect the presence of rat meat