Development of a new set of reference genes for normalization of real-time RT-PCR data of porcine backfat and longissimus dorsi muscle, and evaluation with PPARGC1A
نویسندگان
چکیده
BACKGROUND An essential part of using real-time RT-PCR is that expression results have to be normalized before any conclusions can be drawn. This can be done by using one or multiple, validated reference genes, depending on the desired accuracy of the results. In the pig however, very little information is available on the expression stability of reference genes. The aim of this study was therefore to develop a new set of reference genes which can be used for normalization of mRNA expression data of genes expressed in porcine backfat and longissimus dorsi muscle, both representing an economically important part of a pig's carcass. Because of its multiple functions in fat metabolism and muscle fibre type composition, peroxisome proliferative activated receptor gamma coactivator 1alpha (PPARGC1A) is a very interesting candidate gene for meat quality, and was an ideal gene to evaluate our developed set of reference genes for normalization of mRNA expression data of both tissue types. RESULTS The mRNA expression stability of 10 reference genes was determined. The expression of RPL13A and SDHA appeared to be highly unstable. After normalization to the geometric mean of the three most stably expressed reference genes (ACTB, TBP and TOP2B), the results not only showed that the mRNA expression of PPARGC1A was significantly higher in each of the longissimus dorsi muscle samples than in backfat (P < 0.05), but also that the expression was significantly higher in the most cranial of the three muscle samples (P < 0.05). CONCLUSION This study provides a new set of reference genes (ACTB, TBP and TOP2B) suitable for normalization of real-time RT-PCR data of backfat and longissimus dorsi muscle in the pig. The obtained PPARGC1A expression results, after application of this set of reference genes, are a first step in unravelling the PPARGC1A expression pattern in the pig and provide a basis for possible selection towards improved meat quality while maintaining a lean carcass.
منابع مشابه
Selection of suitable reference genes for real-time PCR studies of early developmental stages of sturgeons
In quantitative real-time PCR, the mRNA level can be quantified in relative terms based on the expression ratio of mRNAs of the target gene and an internal reference gene. Since, an internal standard should be expressed at a constant level among different tissues of an organism at all stages of development, and should be unaffected by the experimental treatment, the stability of different refer...
متن کاملValidation of Reference Genes for Real Time PCR Normalization in Milk Somatic Cells of Holstein Dairy Cattle
Real time-qPCR is the most reliable method for evaluation of mRNA expression levels. However, to obtain accurate results, selection of suitable reference genes is necessary for normalizing the real-time qPCR data. The aim of this research was to validate the expression stability of three potential reference genes (ACTB, GAPDH and UXT) in milk somatic cells of Holstein dairy cattle under differe...
متن کاملThe Expression of Myogenin and Myostatin Genes in Baluchi Sheep
Myogenin gene (MYoG) affects the synthesis of muscle myofibrillar growth and increase of meat production. The myostatin (MSTN) gene is identified as a specific negative regulator of skeletal muscle growth. Reduction of the expression level of MSTN throughmutation in the sequence of this gene leads to an increase of myogenesis and regeneration of muscle cells during the postnatal growing period ...
متن کاملReference Gene Selection in Adipose and Muscle Tissues of Fat-tailed Lori-Bakhtiari Lambs
BACKGROUND: Fat-tailed sheep breeds have a unique ability to tolerate periods of negative energy balance due to seasonal changes in feed availability. This ability is attributed to presence of fat-tail as a body energy reserve, however the exact underlying mechanisms controlling the response of adipose tissue depots to variations in energy balance in fat-tailed breeds a...
متن کاملA Real-Time RT-PCR Assay for Genotyping of Rotavirus Strains
Background: Human rotavirus (HRV) is the causative agent of severe gastroenteritis in children and responsible for two million hospitalizations and more than a half-million deaths annually. Sequence characteristics of the gene segments encoding the VP7 and VP4 proteins are used for the genotype classification of rotavirus. A wide variety of molecular methods are available, mainly based on rever...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- BMC Biotechnology
دوره 6 شماره
صفحات -
تاریخ انتشار 2006