Abstract |
Process enabling simultaneous detection of two transgenes namely human serum albumin (HAS) and bar genes using a multiplex polymerase chain reaction (MPCR) utilizing a combination of novel primers and polymerase chain reaction (PCR) protocol in transgenic wheat". The present invention comprises a process enabling simultaneous detection of two transgenes i.e. human serum albumin (HAS) gene responsible for human blood serum protein production as biopharmaceutical product and bar gene imparting resistance against herbicide phosphimothrecin in transgenic wheat using a Multiplex Polymerase Chain Reaction (MPCR). The process comprises (i) Designing of the primer sequence for human serum albumin (HAS) and bar gene (ii) Preparation of polymerase chain reaction mixture (iii) Amplification of target sequence with three temperatures multiplex polymerase chain reaction (iv) Separation of amplified fragments on 3% agarose gel and staining with ethidium bromide. The process developed in the present invention is much more economical, rapid and precise than the traditional polymerase chain reaction. Bar gene is used as the most common selectable marker sequence to transform several commercialized transgenic crops. Multiplex polymerase chain reaction process can confirm that a plant is genetically modified as bar gene along with human serum albumin (HAS) gene can be detected in single experiment, thus the process is much more reliable also for detection/screening of transgenic crops because the transgenic wheat for human serum protein production and herbicide tolerance targeted in this invention has been imported in India will be commercially grown in the country and other countries as well. It has become all the more important to develop reliable, efficient and precise protocols for detection of the transgenes incorporated in wheat i.e. human serum albumin (HAS) gene and bar gene to meet the regulatory requirement and to solve the legal disputes in future if they arise. |