Running in his penultimate race of his illustrious career, Bolt eased past the finishing line as he helped the defending champions Jamaica win the second heat in 37.95 seconds, their season's best, to enter the final round with third best time.
The Jamaican quartet of Tyquendo Tracey, Julian Forte, Micheal Campbell and Bolt led the eight-team second heat from beginning to the end.
Experienced Yohan Blake, who finished fourth in the men's 100m final and failed to qualify for the final round in 200m, did not start in the heats but Bolt later said that he will return in the final race.
The top three from each of the two heats and next two fastest finishers from the remaining qualify for the final round.
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The United States team of Mike Rodgers, Justin Gatlin, Bee Jay Lee and Christian Coleman won the first heat in 37.70 seconds, the world leading time this season, to clock the overall best time going into the final round.
Great Britain clocked the second best time of 37.76, while France were fourth with their season's best of 38.03.
Bolt, who has anchored the gold-winning Jamaican team in both 2015 World Championships and Rio Games, will be seeking his 20th global title and would want to hang up his boots with a yellow metal in his final race of his glittering career.
He lost the 100m dash last Saturday in his final individual race, settling for a bronze.
Widely considered as the greatest sprinter of all time, Bolt has won eight Olympic and 11 World Championships gold medals beginning with a 100m and 200m double at the 2008 Beijing Olympics.
Since then he has dominated the shorter two sprint events like none in athletics history. His Jamaican team had also won the 4x100m relay gold in Beijing but it had to return the medal after one member of the quartet -- Nesta Carter -- tested positive for a prohibited substance.
Whenever the giant screen inside the stadium showed him doing the baton exchange practice with his Jamaican team-mates in preparation for the 4x100m relay race at the warm-up arena, the capacity crowd cheered loudly.
The showman, as usual, entertained the crowd as he acknowledged the cheers as he entered the stadium for the relay heats. He did his routine marking of his starting position and did a few run-downs imitating baton exchange.
"There are no words to describe how I am feeling. I get so much support from the crowd. I appreciate that a lot. All the energy I am getting from the crowd. It is brilliant," Bolt said after the race.
"The young runners in our team -- it is just about executing and coming through the race for them. We have been training that relay, there were some camps. Still there were some mistakes. Yohan Blake is coming in for the final is definitely good."
Olympic champions United States won the first heat in a world leading time of 41.84 seconds, even in the absence of individual 100m champion Torie Bowie.
Great Britain finished second in that heat -- and also second overall -- with 41.93, the third fastest time ever produced by a British quartet.
Defending champions Jamaica (42.50) finished second behind Germany (42.34) in the second heat.
American runner Allyson Felix later said that Bowie will return in the final round.
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