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

Authors: AJAI TRIPATHI, KRISHNA M SINGH, MAKRAND N. VAZE, UMED V. RAMANI, DHARAMSI N. RANK, CHAITANYA G. JOSHI

Abstract: In the present study, antisense primers were designed from 7 differentially displayed expressed sequence tags (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 (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 (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, \beta-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: ESTs, RACE, full length, cDNA, rRNA

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