The Panthers are hoping to secure back-to-back wins, while the visitors will be aiming for their first points in the qualifying campaign.
Match preview
Gabon rebounded from a 4-1 drubbing by Morocco in the opening game of their AFCON qualifying series by securing a 2-0 victory over Central African Republic in their second match a couple of days later.
Pierre-Emerick Aubameyang converted a second penalty kick in as many AFCON qualifying games, putting the Panthers ahead, and Shavy Babicka doubled the lead in the 40th minute to seal the win.
Gabon climbed to second place in Group B with three points from a possible six, level with Central African Republic, who are behind due to an inferior head-to-head record.
The hosts enter Friday's clash seeking to claim a victory in the first of two successive games against Lesotho, a side they have never lost against, winning three and drawing one of four encounters.
All the meetings between both sides have been in qualifying matches - two each for the World Cup and AFCON - and the Panthers won both times they have hosted this fixture.
Lesotho kicked off their AFCON qualifying campaign with a 3-1 defeat to Central African Republic in the first round of qualifiers and were forced to react in their second game against Morocco.
However, the Southern African outfit, who played on Moroccan soil due to the lack of an international-standard stadium back home, succumbed late in the game, conceding a 93rd-minute goal to lose 1-0.
Currently bottom of the group, Leslie Notsi's side will hope to pick up their first set of points when they travel to Gabon on Friday, as they are now on a six-match winless streak, which includes a disappointing campaign at the COSAFA Cup in July.
The visitors secured a 2-0 win away against Zimbabwe in June in a World Cup qualifying match, a performance and result they will hope to replicate against Gabon on Friday.
Gabon Africa Cup of Nations Qualifying form:
L
W
Gabon form (all competitions):
W
L
D
L
L
W
Lesotho Africa Cup of Nations Qualifying form:
L
L
Lesotho form (all competitions):
L
D
L
L
L
L
Team News
A front three of Aubameyang, Babicka and Denis Bouanga is expected to lead the line for Gabon, with Aubameyang hoping to make it three straight matches with a goal.
A midfield trio of Eric Bocoum, Guelor Kanga and Medwin Biteghe started in the last match and will aim to retain their places in the starting lineup.
Meanwhile, Lesotho have called up five new players to their 31-man provisional squad, including 19-year-old Paseka Maile, who made his senior team debut at the COSAFA Cup earlier this year.
Sera Motebang, who has scored the Crocodiles' only goal in the AFCON qualifiers so far, is expected to lead the attack once again.
Gabon possible starting lineup:
Mbaba; Ekomie, Appindangoye, Moucketou-Moussounda, Oyono; Biteghe, Kanga, Nziengui; Bouanga, Aubameyang, Babicka
Lesotho possible starting lineup:
Moerane; Makhele, Malane, Mokokoane, Matlabe; Fothoane, Toloane, Lebokollane, Mokhachane; Makateng, Thabantso