Skin is one of the largest organ of the human body which accounts for almost 15% body weight. The skin also known as first line of defence act as a protective barrier against water loss from the body and external environmental stress like UV radiations. The human skin run along the entire stretch of body. This stretch mainly comprises of layers as Epidermis, Dermis and Sub cutaneous hypodermal layer. The epidermal region is comprised of the cells like Keratinocytes, Melanocytes, Merkel cells and Langerhans cells. Keratinocytes present in epidermal region undergo series of differentiation and sequential processes to give four distinct layer of the epidermis. These layers are Stratum Corneum, Stratum Lucidium, Stratum Granulosum, Stratum Spinosum and Stratum Besale. The Keratinocytes produces growth factors like SCF, ET-1, ?MSH which bind to the receptors present on the plasma membrane of the melanocytes, once the factors are bound to the receptors, the melanocytes start producing the pigment Melanin a dark colour pigment that impart skin it’s colour and it serves as protection against UV radiations
Mast cell growth factor commonly known as the Stem Cell Factor is the multifunctional cytokine that is involved in the melanogenesis, hematopoiesis and gametogenesis. It triggers its biological effect by binding to the c-kit (the SCF receptor). It induces melanogenesis by the interaction of keratinocytes and melanocytes. The presence of Exon 6 which encodes proteolytic cleavage producing soluble form, allows the Scf to exist in two forms that are soluble form(inclusion) and transmembrane(exclusion). Both the forms are biologically active. Although the stem cell factor has a wide area of functionality but cost of Scf production from cell line is quite difficult. The rDNA technology has given the solution to this problem. As with the use of rDNA technology we are able to produce various proteins that are valuable in various sectors. The recombinant technology has taken advantage of the bacterial model to produce the human proteins as normally it is very difficult to isolate the human protein.
The bacterial model that we choose is Escherichia Coli as it is most suitable bacterial for expression system, it has high rates of replication ,cost effective and it can produce various proteins and metabolites.
In this project we are incorporate SCF into the cloning vector and will carry out its protein expression using E. Coli and once expressed we will purify the protein.