THE TWO-HYBRID SYSTEM TO EVALUATE PROTEIN-PROTEIN

The Two-Hybrid System to Evaluate Protein-Protein

Abstract

Environmental conditions can have a major impact on the developmental progression in animals. The development of Caenorhabditis elegan known as C.elegans consists of six stages: one embryonic stage, four larval stages ending up with a molt, and one adult stage. The development timing has regulated at every stage and produces cell in each developmental period. The hetero chronic genes are found in the developmental timing in C.elegans, which controls the specificity of cell selection in somatic tissues. As per hypothesis, the protein produced by the lin-66.22 and PF10-28-AL plays an important role in controlling the developmental timing, and these genes cause either premature or retarded developmental phenotypes. The study determines the role of lin-66.22 and PF10-28-AL proteins in the developmental timing of C.elegans. It was observed that lin-66 single mutants displayed the retarded developmental stages. Due to the time limitation, we had prepared the liquid culture by sterling the TA yeast library transformation but the bait plasmid could not be determined. Because of some technical problem with the gel electrophoresis, and the DNA sequencing was not done for this experiment due to the lack of results.The Two-Hybrid System to Evaluate Protein-Protein

Introduction

C. elegan had used as sample for this study. The reason for choosing this organism was to find out how proteins help in the developmental stages of C.elegans. Caenorhabditis elegan belong to the family of Rhabditidae. It is a small transparent worm, of about a millimeter long; it recurs after every three days. This organism has six developmental stages; embryonic stage, four larval stages, molt stage, and last one adult stage. The hetero chronic genes are important in governing, and controlling the timing and sequence of the larval stages of C.elegans.

Literature review

Two mutations, unc-86 and lin-4, changed the usual invariant progress of cell clones in the round worm like C.elegans. The different cell clones of these mutants became stuck at some point in their development and repeat the patterns of cell divisions. Lin-4 mutant worms are unable to otkladyat eggs, other mutations lead to the inability to lay eggs. Mutants unable to lay eggs showed that the mutation lin-14, lin-28 and lin-29 are associated with abnormal progress of cell clones. These mutations lead to two tipamneregulyarnoy progression of cell cloning in which specific ontogenetic processes take place. In wild animals, these processes occur later than the normal time. This is the reason that new mutations have identified as heterochronic (Banerjee, 2005).

The most interesting gene is lin-14, in which different alleles have protivovolozhnye phenotypes of premature or delayed progression of cell fates. Protein expression of lin-14 mutants with abnormal function has preserved at all stages of larval development has observed even in adult worms. Lin-14 antagonistically has regulated by lin-28 and lin-4.

Yeast Strain L40, with a genotype of Mata ade2 his3 leu2 trp1 LYS: lexA-HIS3 URA3: lexA-lacZ can be used as a host for determining protein- protein interactions. Yeast strain L40 contains LexZ which produce galactosidase, and HIS3 reinitiates a ...