Full length cDNA synthesis of differentially displayed ESTs during lactation in the Indian buffalo (Bubalus bubalis)

Authors : Ajai K TRIPATHI, Krishna M SINGH, Makrand N VAZE, Umed V RAMANI

Pages : 107-113

Doi:10.3906/sag-1205-21

View : 19 | Download : 10

Publication Date : 0000-00-00

Article Type : Research Paper

Abstract :In the present study, antisense primers were designed from 7 differentially displayed expressed sequence tags insert ignore into journalissuearticles values(ESTs); previously identified at our laboratory. Total RNA was isolated from the mammary gland tissue of lactating buffalo to carry out 5` rapid amplification cDNA ends insert ignore into journalissuearticles values(RACE); PCR. 5` RACE PCR amplicons, ranging from 161 bp to 336 bp, were cloned and sequenced. These 5` RACE sequences were analyzed using the blastn insert ignore into journalissuearticles values(Megablast); bioinformatics tool available from NCBI. Of these 7, only one 5` mRNA sequence did not show a match in the available database and another 1 failed to show a match with any specific gene due to the unavailability of buffalo genome information. Two other 5` mRNA sequences showed 82% to 100% homology with the 18S rRNA gene and the remaining 3 showed homology to the Talin-1, b-casein, and 28S rRNA genes. Full length cDNA were synthesized using primers designed from 5` mRNA sequences and ranged from 345 bp to 1030 bp. Finally, we conclude that the ESTs identified may act as markers for lactogenesis in other animals.
Keywords : Key words ESTs, RACE, full length, cDNA, rRNA