One of the most important parameter in elongation of cotton fibre is the biosynthesis of ethylene. EIN3 and EIL1 (Ethylene Insensitive3/Ethylene Insensitive 3) are the main transcriptional factors that are specifically playing an important role in signal transduction in cotton. Expression analysis of the EIN3/EIL1 genes involved in development of cotton fibre investigated out through RT-qPCR. Further, STRINGTIE is a tool that is use for measuring the different levels of expression. ClustalW method is a method that is used to determine the sequences of amino acids of EIN3/EIL1 genes by aligning by using the sequences. KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways of the EIN3/EIL1 genes is also bioinformatics tools that is functionally use for classification and characterization of genes at functional level with using the principles of functional enrichment analysis. Biological database such as STRING is designed for to construct the specific PPI networks of EIN3/EIL1 due to known PPIs and analysis to check interactions among protein. Bioinformatics tool such as BLASTP use for identification of transcription factors of EIN3/EIL1genes. Biological database such as Plant CARE is especially used to analyze the cis-elements in specific sequences of promoters of EIN3/EIL1 that are especially involved in transcription of genes.