Cloning and Molecular Characterization of Lectin Rgene From Garlic (Allium Sativum L.)

Abstract

Researchers have shown that insect pests' viz. planthoppers and aphid's damages many crops and also transmit many viral diseases. The detrimental mechanism of action of plant lectins for pests by binding to the larval brush border and peri trophic membrane, through disruption of microvilli, by change in the gut membrane environment or through disruption of digestive enzyme in well known. Although lectins are considered to be toxic for mammals and results in loss of appetite, vomiting, diarrhoea, decreased body weight and eventually death, but not all lectins are toxic for mammals. Thus, in the present study attempt was made to isolate and clone and characterize Allium sativum lectin gene. The gene was isolated using gene specific primers designed by FastPCR, which resulted in a 1030bp sequence. The band was eluted and gene was ligated with the A/T cloning vector which was followed by confirmation using plasmid DNA isolation, colony PCR and Restriction digestion approaches. The positive results were subjected to sequencing and were further subjected to insilica analysis. The results presented that the query sequence similar to many other lectins with accession numbers M85174, M85175, M85176 etc. The sequence was also compared for the presence of mannose binding region because mannose binding lectins (M:MBL ), esp. garlic lectins, are although effective for controlling insect pests, but are nontoxic to mammals, birds and are thus believed to be the safest agglutinin for diet. The results showed a higher similarity of query sequence with other lectins in this regard. Many lectin genes have been isolated but much more needs to isolated, the mode of action can be improved and new applications of lectins need to be found. The attempt to isolate, incorporate Garlic lectin genes or related genes with similar characteristics and development of more resistant plants against pests has a good potential to satisfy~eman or production by increasing the crop yield.