Abstract:
So far, many promoters have been isolated and used to drive the expression of foreign gene in the mammary gland. However, none of these promoters were specific and the expression in the mammary gland tissue was associated with expression in other tissues. The need for discriminating sequences, which can drive the expression of foreign gene specifically in the mammary gland, is obvious. This study is intended to clone specific regulatory elements of 11 casein gene, which may drive the expression of every gene specifically in the mammary gland. The isolation and purification procedure of the genome goat from the mammary gland and the isolation and purification of Lambda vector was performed.
The right and left arms were of the phage were also purified and packaged with packagene. A goat genome library consisting of 2. 1 X 105 phages was constructed and screened by using in situ hybridization technique. Following the procedure of Sambrook et al. (1989) we performed the in-situ hybridization technique. The probe was prepared through Polymerase chain reaction (PCR) using the goat genome as template. The probe was used in the in-situ hybridization technique to screen the library after cloning it into puc 18 vector. Sall and BamHI were used for the analysis of the recombinant phage. The sal 1 fragments were cloned in pGEM5z vector. The partial sequencing of the 4.5kb long Sal 1 fragment cloned in pGEMbz was performed following the procedure of Sambrook et al. (1989). In this study we suggest that the goat 11 casein promoter may be very strong and tissue specific promoter, which may contribute in the progress and development of the mammary gland expression for the production of valuable proteins in the milk.