Abstract

CLONING AND EXPRESSION OF NOVEL LINGO-1 GENE FROM BUBALUS BUBALIS

Imteyaz Qamar*, Nagendra Singh, Sahar Waseem, Prapti Prakash, Veerendra Singh Nagoria and Mohammad Faiz Ahmad

ABSTRACT

In this study, we have identified, cloned and expressed full length Lingo-1 gene from water buffalo Bubalus bubalis testis cDNA. LINGO-1 gene is well studied in various animal models. But it is a novel protein to be studied in Bubalus bubalis. Previous studies by other groups uncovered several transcripts representing known and novel genes tagged with consensus of 33.15 using cDNA from the testis, ovary, spleen, kidney, heart, liver and lung of water buffalo Bubalus bubalis. Among those transcripts one was found to represent the partial cDNA sequence of LRRN6A. Studies of transcripts using Minisatellite-Associated Sequence Amplification (MASA) showed thepresence of LRRN6A in testis of water buffalo Bubalus bubalis. The cloning strategy involved amplification of whole length by using five different pairs of degenerate primers. The amplified fragments were sequenced and aligned to obtain the whole gene sequence. Full length gene was amplified using gene specific primers and successfully cloned and expressed in bacteria. The gene was found to have significant homology to the LINGO-1 gene from Bos taurus. Further transcript analysis from different organs of Bubalus bubalis indicates an organ specific expression of LINGO-1 gene.

Keywords: Bubalus bubalis, Bos Taurus, cDNA, LRRN6A.