Gene structure analysis of caprine and ovine Mannose Binding Lectin (MBL)

ISSN 1594-4077

Vol.8

-

2OO9

Supplement

2

www.aspajournal.

it

ITnLIAN

JOURNAL

OF

ANIMAL

ScrENcE

Journal and Official

Organ

of

the

Scientific

Association

of Animal

Production (ASPA)

THE

ASPA

(b

_ulJ

EdizioniAvenue media'

Milano -Bologna

Gene

structure

_anarysis

_{of caprine}

_and

ovine

Mannose

_{Binàing Lectin}

_rrrreLl-.

Gianfranco

_cosenza,

_{Arfredo pauciuilo, Letizia corimoro,}

Andrea

Mancusi,

_Anna

_Lidia

_Annunziata,

Daniela

_Gallo,

_Luigi

_Ramunno

Dipar[imento dÌ scienze del suolo, della Pianta, dellAmbiente _e delle produzioni Animali Università

di

Napoli ,,Federico

_II",

_Italy

coTesponcÌing auihor" _{Luigr Ramunno Diparrimento}

_di

scienze del suo10. della pianta. deilAmbiente e

delle Produzioni _{-{nrmah universrtà di Napoli "Federico}

II".

\iia

u""..Ja

100. 80055 portici (NA). Italv

- Tel. +39 0S1 25S900.1 - Fax: +39 OSf ;Z.ZÉSO _ _Email:

_{.u_ro.roO,r.riou:t}

ABSTRACT

The Mannose Binding

Lectin

(MBL)

_is

_an

_importaat

_factor

of

natural immunity

_{r.vhich acts as a}

primordial

_{antibodr', capable}

_{to acti'ate the}

complement and to

link

to _{some sugars (such as}

man-nose) present on the surface of some bacteria as Brucelia.

_L

_thu

_p."*rt

_{stud1.. we report}

the

partial

structure of the gene coding for the

MBL

in

sheep and goat as

weil

as the

a,alysis

of

their

promot-ers' By using as template genomic DNA from leukoc5.-tes _{obtained from}

_indi'idual

blood samples we

sequenced from -961

nt

of the promoter to the 397th

nt

of the 4,h exon of fbur sheep (EMBLAcc. _no. 4M933378) _{and from -97g}

_nt

_{to the 471th}

_nt

of the 4th exon of to,,,,

gort.

(EMBL _{Acc. no. AMg33377),} for a

total

of _{4467 bp and 4515 bp, respectivellr 85.'comparing}

rrr" _{rr"o*oìogous sequence of the bo'ine}

MBL

gene

(EMBLAcc

_no

_{NC007327),itrvas}

possible to determine the size of the exons,

similar in

both investigated _{species:200 bp (exon 1), 117 Lp (exon 2). and 69 bp (exon}

3). _{The comparison}

_ofthe

sequences betrveen goat and sheep shor.r,s an homoiogl-

of

_{g2.2Tc.The differences are due essentialÌy}

to insertions/deletions at

intronic

level: the sheep presents

r-ith

respect to the goat a deletion at the

1"'intron

of 11

_{nt' t*o}

_{insertrons at the 2"d intron.}

respectivelv of 29 and 41

nt,

and at the 3.d

intron

a stretch _{of eighr Guanrae rn tbe eoat r.s eiohr}

_l

u,as _{3e5 bp us 385}

_{r,"*"-,lr-"r"}

o'J::'Jl?:i.1:ffi,'Jr'il;tr,Jil#?flil1:T"lJn,T[:l;

goatandsheep Forbothspecies.allthesplicejunftionsfoilou-the.oo.-o.,role5,GT/3,AG.TheoRF

(open _{Reading Frame) encodes for}

249 amino _{acid residues. u,hereas the Ìeader peptide is}

composed by 19 amino acids The startrng codon (ATG, Methionine) _{is located between the 10to and the}

12tr nt o{.

the 1"i exon' _{in'hile the stop codon (TGA) bet'u,'een the}

B72ih _{a..d the 374th}

_nt

_{of the 4th exon. \\4rile}

from

the _{comparison of} the _{exonic sequences, 23 sNPs were identified,}

10

oirrhich

give _{rise to an amino} acidic change The sequence analysis of the promoter _region

_in

_{sheep shou,s the}

presence of the

f.oi-lowing _{putative binding sites for transcription}

factors: TATA_box (ATA_$) _{and CCA*A.T_box (CCA-AAT)}

in

position _{-49 and -140 nt, respectivelv'.}

Furthermore,

_{;;;r*;.,o..iloi}

_.".poo=ive

erements (GR_EJ

were identified:

in

_{position -765 (AGATCAGA) and -971}

_{(ACiGATcD. irre}

promorer anarysis for goat was found to be analogous to

that

ofsheep but

it

shows the disappearrrr."

_{orrrr. reg,ìatory}

_{site GRE}

(ACTGATCT).

_Further

studies need to be performed

in

order to verifi, whether the markers identified

in

_{the present study can be associated to}

_ìhe

resistance to pathogens such as Brucella _{melitensis bv} carrying out suitable test for antimicrobial actir_itr,.

196

IraL.J.Ar,irM.Scr. _{vor. B (Suecr,}

_2),